• Title/Summary/Keyword: Two-stage fermentation

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Characteristics of Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) Production by Ralstonia eutropha NCIMB 11599 and ATCC 17699

  • Song, Jae-Yong;Kim, Beom-Soo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.6
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    • pp.603-606
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    • 2005
  • Ralstonia eutropha NCIMB 11599 and ATCC 17699 were grown, and their productions of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] compared. In flask cultures of R. eutropha NCIMB 11599, cell concentration, P(3HB-co-4HB) concentration and polymer content decreased considerably with increases in the ${\gamma}-butyrolactone$ concentration, and the 4HB fraction was also very low (maximum 1.74 mol%). In fed-batch cultures of R. eutropha NCIMB 11599, glucose and ${\gamma}-butyrolactone$ were fed as the carbon sources, under a phosphate limitation strategy. When glucose was fed as the sole carbon source, with its concentration controlled using an on-line glucose analyzer, 86% of the P(3HB) homopolymer was obtained from 201g/L of cells. In a two-stage fed-batch culture, where the cell concentration was increased to 104g/L, with glucose fed in the first step and constant feeding of ${\gamma}-butyrolactone$, at 6g/h, in the second, final cell concentration at 67h was 106g/L, with a polymer content of 82%, while the 4HB fraction was only 0.7mol%. When the same feeding strategy was applied to the fedbatch culture of R. eutropha ATCC 17699, where the cell concentration was increased to 42 g/L, by feeding fructose in the first step and ${\gamma}-butyrolactone$ (1.5g/h) in the second, the final cell concentration, polymer content and 4HB fraction at 74h were 51g/L, 35% and 32 mol%, respectively. In summary, R. eutropha ATCC 17699 was better than R. eutropha NCIMB 11599 in terms of P(3HB-co-4HB) production with various 4HB fractions.

Biohydrogen production from engineered microalgae Chlamydomonas reinhardtii

  • Kose, Ayse;Oncel, Suphi S.
    • Advances in Energy Research
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    • v.2 no.1
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    • pp.1-9
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    • 2014
  • The green microalgae Chlamydomonas reinhardtti is well-known specie in the terms of $H_2$ production by photo fermentation and has been studying for a long time. Although the $H_2$ production yield is promising; there are some bottlenecks to enhance the yield and efficiency to focus on a well-designed, sustainable production and also scaling up for further studies. D1 protein of photosystem II (PSII) plays an important role in photosystem damage repair and related to $H_2$ production. Because Chlamydomonas is the model algae and the genetic basis is well-studied; metabolic engineering tools are intended to use for enhanced production. Mutations are focused on D1 protein which aims long-lasting hydrogen production by blocking the PSII repair system thus $O_2$ sensitive hydrogenases catalysis hydrogen production for a longer period of time under anaerobic and sulfur deprived conditions. Chlamydomonas CC124 as control strain and D1 mutant strains(D240, D239-40 and D240-41)are cultured photomixotrophically at $80{\mu}mol\;photons\;m^{-2}s^{-1}$, by two sides. Cells are grown in TAP medium as aerobic stage for culture growth; in logarithmic phase cells are transferred from aerobic to an anaerobic and sulfur deprived TAP- S medium and 12 mg/L initial chlorophyll content for $H_2$ production which is monitored by the water columns and later detected by Gas Chromatography. Total produced hydrogen was $82{\pm}10$, $180{\pm}20$, $196{\pm}20$, $290{\pm}30mL$ for CC124, D240, D239-40, D240-41, respectively. $H_2$ production rates for mutant strains was $1.3{\pm}0.5mL/L.h$ meanwhile CC124 showed 2-3 fold lower rate as $0.57{\pm}0.2mL/L.h$. Hydrogen production period was $5{\pm}2days$ for CC124 and mutants showed a longer production time for $9{\pm}2days$. It is seen from the results that $H_2$ productions for mutant strains have a significant effect in terms of productivity, yield and production time.

Effects of Monensin Administation on Mammary Function in Late Lactating Crossbred Holstein Cattle

  • Thammacharoen, S.;Chanpongsang, S.;Chaiyabutr, N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.12
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    • pp.1712-1718
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    • 2001
  • An experiment was carried out to study the effect of monensin administration on mammary functions in crossbred Holstein cattle. Fourteen non-pregnant late lactating crossbred Holstein cattle, approximately 270 days postpartum, were selected for the experiment. They were divided into two groups of 7 animals each. Seven animals in the treated group were given sodium monensin orally in a slow-release capsule. Animals in both control and treated groups were fed the similar diet to maintain milk production and body score at 2.5. Rice straw was fed as a source of dietary fiber throughout the experimental period. After monensin administration, a significant increase in the molar percent of ruminal propionate (p<0.05) and a significant decrease in the molar percent of ruminal acetate (p<0.05) were apparent in comparison to the pretreated period. The ratio of acetate to propionate concentration decreased significantly after monensin administration (p<0.05), while it was maintained at the similar level throughout the period of experiment in the control group. Monensin did not affect the molar percent of ruminal butyrate and valerate. The concentration of milk allantoin between the control group and monensin treated group was not different. An excretion rate of allantoin in milk decreased in animals treated with monensin (p<0.05). Mammary blood flow did not show significant difference between control and monensin treated groups. The plasma glucose concentration, arteriovenous concentration difference and mammary gland uptake of glucose remained constant in both groups. Milk yield of the later stage of lactation in the control group declined during lactation advance while a tendency to increase in the milk yield was apparent after 21 days monensin administration. Milk compositions for concentration of lactose, fat and protein in both control group and monensin treated group did not change throughout the experimental periods. From these results, it can be concluded that the action of monensin could affect the ruminal fermentation pattern. Monensin could not increase milk yield in the late lactating period.

Effects of Addition Level and Chemical Type of Propionate Precursors in Dicarboxylic Acid Pathway on Fermentation Characteristics and Methane Production by Rumen Microbes In vitro

  • Li, X.Z.;Yan, C.G.;Choi, S.H.;Long, R.J.;Jin, G.L.;Song, Man K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.1
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    • pp.82-89
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    • 2009
  • Two in vitro experiments were conducted to examine the effects of propionate precursors in the dicarboxylic acid pathway on ruminal fermentatation characteristics, $CH_4$ production and degradation of feed by rumen microbes. Fumarate or malate as sodium salts (Exp. 1) or acid type (Exp. 2) were added to the culture solution (150 ml, 50% strained rumen fluid and 50% artificial saliva) to achieve final concentrations of 0, 8, 16 and 24 mM, and incubated anaerobically for 0, 1, 3, 6, 9 and 12 h at $39^{\circ}C$. For both experiments, two grams of feed consisting of 70% concentrate and 30% ground alfalfa (DM basis) were prepared in a nylon bag, and were placed in a bottle containing the culture solution. Addition of fumarate or malate in both sodium salt and acid form increased (p<0.0001) pH of culture solution at 3, 6, 9 and 12 h incubations. The pH (p<0.0001) and total volatile fatty acids (VFA, p<0.05) were enhanced by these precursors as sodium salt at 3, 6 and 9 h incubations, and pH (p<0.001) and total VFA (p<0.01) from fumarate or malate in acid form were enhanced at a late stage of fermentation (9 h and 12 h) as the addition level increased. pH was higher (p<0.001) for fumarate than for malate as sodium salt at 3 h and 6 h incubations. Propionate ($C_3$) proportion was increased (p<0.0001) but those of $C_2$ (p<0.05) and $C_4$ (p<0.01 - p<0.001) were reduced by the addition of sodium salt precursors from 3 h to 12 incubation times while both precursors in acid form enhanced (p<0.011 - p<0.0001) proportion of $C_3$ from 6h but reduced (p<0.018 - p<0.0005) $C_4$ proportion at incubation times of 1, 3, 9 and 12 h. Proportion of $C_3$ was increased (p<0.05 - p<0.0001) at all incubation times by both precursors as sodium salt while that of $C_3$ was increased (p<0.001) from 6h but $C_4$ proportion was decreased by both precursors in acid form as the addition level increased. Proportion of $C_3$ was higher (p<0.01 - p<0.001) for fumarate than malate as sodium salt from 6 h incubation but was higher for malate than fumarate in acid form at 9 h (p<0.05) and 12 h (p<0.01) incubation times. Increased levels (16 and 24 mM) of fumarate or malate as sodium salt (p<0.017) and both precursors in acid form (p<0.028) increased the total gas production, but no differences were found between precursors in both chemical types. Propionate precursors in both chemical types clearly reduced (p<0.0001 - p<0.0002) $CH_4$ production, and the reduction (p<0.001 - p<0.0001) was dose dependent as the addition level of precursors increased. The $CH_4$ generated was smaller (p<0.01 - p<0.0001) for fumarate than for malate in both chemical types. Addition of fumarate or malate as sodium type reduced (p<0.004) dry matter degradation while both precursors in both chemical types slightly increased neutral detergent fiber degradability of feed in the nylon bag.

Treatment of highly concentrated organic wastewater by high efficiency $UV/TiO_{2}$ photocatalytic system (고효율 자외선/광촉매 시스템을 이용만 고농도 유기성 폐수처리)

  • Kim, Jung-Kon;Jung, Hyo-Ki;Son, Joo-Young;Kim, Si-Wouk
    • KSBB Journal
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    • v.23 no.1
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    • pp.83-89
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    • 2008
  • Food wastewater derived from the three-stage methane fermentation system developed in this lab contained high concentration organic substances. The organic wastewater should be treated through advanced wastewater treatment system to satisfy the "Permissible Pollutant Discharge Standard of Korea". In order to treat the organic wastewater efficiently, several optimum operation conditions of a modified $UV/TiO_{2}$ photocatalytic system have been investigated. In the first process, wastewater was pre-treated with $FeCl_{3}$. The optimum pH and coagulant concentration were 4.0 and 2000mg/L, respectively. Through this process, 52.6% of CODcr was removed. The second process was $UV-TiO_{2}$ photocatalytic reaction. The optimum operation conditions for the system were as follows: UV lamp wavelength, 254 nm; wastewater temperature, $40^{\circ}C$; pH 8.0; and air flow rate, 40L/min, respectively. Through the above two combined processes, 69.7% of T-N and 70.9% of CODcr contained in the wastewater were removed.

Treatment of High Organic Wastewater Using Ecological Water Treatment System (생태학적 수처리 시스템을 이용한 고농도 유기성 폐수처리)

  • 조재훈;김중곤;김준휘;윤성명;이정섭;김시욱
    • Korean Journal of Microbiology
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    • v.37 no.4
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    • pp.317-324
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    • 2001
  • We have previously developed three stage methane fermentation system capable of digesting food wastes effectively and then releasing high organic wastewater as a final product. In this study, we tried to devise an ecological water treatment system, which can efficiently remove the nitrogen and phosphorus contained in the organic wastewater. The system was made of microbiological filters, algae, and waterfleas. Of two species of alga tested, Selenastrum capricornutum showed higher growth rate and more efficiently removed the nitrogen from the wastewater than by Chlorella sp. In addition, the highest growth rate and the nitrogen removal efficiency could be obtained when high concentrations of $Mg^{2+}\; and\; Ca^{2+}$ were added to the diluted wastewater and the molar ratio of nitrogen to phosphorus was adjusted to 10 : 1. In this study the population relationship between alga and water flea was also examined in a test tube. The initial number of algal cells decreased as the waterflea population increased. However, the number of algal cells gradually increased again when waterflea population decreased partly due to the environmental resistance. From these results, it was believed that the ecological water treatment system could be used for removing the nitrogen and phosphorus from organic wastewater very effectively. Moreover, the waterflea cultured by this system as a final predator could be used as a good foodstuff for fishes.

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Studies on the Induction of Available Mutant of Acetic Acid Bacteria by UV-light Irradiation and NTG Treatment. - The Selection of Mutant Strains and Various Conditions for Acetic Acid Production - (Acetobacter sp.와 그 변이주(變異株)를 이용(利用)한 식초산(食酢酸) 발효(醱酵)에 관한 연구(硏究) - 변이주(變異株)의 선정(選定) 및 생산조건(生酸條件) -)

  • Kim, Chan Jo;Park, Yoon Joong;Lee, Seuk Keun;Oh, Man Jin
    • Korean Journal of Agricultural Science
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    • v.7 no.2
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    • pp.169-175
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    • 1980
  • These studies were conducted to induce the available mutant strains in acetic acid bacteria by the irradiation of UV-light and the treatment of N-methyl-N'-nitro-N'-nitrosoguanidine. 109 strains which were capable of producing acid in the ethanol containing medium were isolated from vinegar and Kimchi collected from the region of Daejeon city. From the collection T-50 strain was identified to have a strong fermentation power and selected as a mother strain for the study. Two mutants were obtained by treating T-50 strain with UV and NTG, and these mutants had a rapid acid production in the initial stage. The study was then made to compare the basic condition for acetic acid production of the mother strain and two mutant strains. The summarized results were as follows; 1. The isolated strain (T-50) was identified as Acetobacter aceti by Bergey's manual and Acetic acid bacteria (Tokyo Univ. press). 2. The selected strain was died completely when the strain was irradiated with 15 W UV-light at a distance of 45 cm for 160 seconds. 3. The mutants such as U-48 and N-67 were rapid in the acetic acid production at the initial stage compare to the mother strain. 4. Acetic acid formation by the shaking culture method was maximized in 2 days culture, and the optimal temperature for acetic acid production was $30^{\circ}C$. 5. Acetic acid was effectively produced by the addition of 0.1% ammonium sulfate as a nitrogen source and was also produced rapidly by the addition of 0.1% glucose.

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Effect of Vinegar Made with Hydroponic-cultured Panax ginseng C. A. Meyer on Body Weight and Lipid Metabolism in High-Fat Diet-Fed Mice (수경재배 인삼으로 제조한 식초가 고지방 식이를 섭취한 마우스의 체중 및 지질대사에 미치는 영향)

  • Oh, Young-Joo;Kwon, Seung-Hyuk;Choi, Kum Boo;Kim, Tae-Seok;Yeo, Ik-Hyun
    • Korean Journal of Food Science and Technology
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    • v.46 no.6
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    • pp.743-749
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    • 2014
  • The aim of this study was to develop hydroponic-cultured ginseng vinegar (HGV) containing ginsenoside Rg2 in order to its anti-obesity and anti-hyperlipidemic effects in C57BL/6J mice. HGV was prepared by two-stage fermentation. The ginsenoside Rg2 contained in acetic acid-fermented HGV increased by 4.0 times compared to that in pre-fermented HGV. To measure the anti-obesity effect of HGV, thirty two mice were randomly divided into four groups: normal diet group (ND), high-fat diet group (HFD), high-fat diet-supplemented with HGV group (HGV), and high-fat diet-supplemented with green tea extract group (GT). Body weight, fat weight, and liver weight decreased in the HGV group. The HGV group also showed lower plasma levels of low-density lipoprotein (LDL)-cholesterol and triglycerides, and higher levels of high-density lipoprotein (HDL)-cholesterol compared to the corresponding levels in the HFD group. Furthermore, there were significant decreases in plasma aspartase aminotransferase (AST) and alanine aminotransferase (ALT) levels in the HGV group compared to the corresponding levels in the HFD group. These results suggest that HGV can be used as an anti-obesity therapeutic agent or functional ingredient.

Effects of Total Mixed Rations on Ruminal Characteristics, Digestibility and Beef Production of Hanwoo Steers (섬유질배합사료 급여가 비육후기 거세한우의 반추위 발효성상, 소화율 및 산육성에 미치는 영향)

  • Kim, K.H.;Kim, K.S.;Lee, S.C.;Oh, Y.G.;Chung, C.S.;Kim, K.J.
    • Journal of Animal Science and Technology
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    • v.45 no.3
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    • pp.387-396
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    • 2003
  • This experiment was carried out to compare the effects of feeding commercial formula feed and rice straw separately (control) versus a total mixed ration (TMR) on productivity of Hanwoo steers in late stage of fattening and on ruminal fermentation characteristics and digestibilities. Ruminal digesta from the cannulated cattle were sampled at 0, 1, 2, 3, 5, 8 hour after feeding. The steers fed TMR consumed 7.4kg per day and there was no difference between feeding systems. Daily weight gain was not significantly (P>0.05) between feeding systems, however, TMR group showed lower daily gain than control group. The amount of feed consumption per kg weight gain was higher in TMR group than control group (10.5kg and 9.7kg, respectively), resulting in a greater efficiency of feed utilization for gain. In the result of appearance rates of quality grade A were 33% higher for TMR group than those in control group. Appearance rates of grade 1 showed 56% and 75% when fed the control and TMR, respectively. Digestibilities of dry matter, crude protein, crude fiber and gross energy for TMR treatment were significantly higher (P<0.01) than those of control. Prior to feeding (0 h) and each subsequent hour, the TMR resulted in higher rumen pH (P<0.05) when compared with control ration. The concentration of NH3-N for TMR treatment maintained at higher level up to 8hr after feeding, especially increased up to 28.2mg/$d\ell$ during 1-2 hour which was two times (P<0.05) more than control. The amount of total VFA showed same trends between feeding systems. However, the ratios of branched chained fatty acid such as iso-butyric acid and iso-valeric acid for TMR treatment were significantly(P<0.01) higher than control for 3-5hr. Results showed that TMR in these trials is effective feeding system for fattening Hanwoo steers in the respect of ruminal characteristics, total tract digestibility and productivities.

A Comprehensive Study on the Forced Aging of Flue-cured Tobacco-Leaves (황색종 잎담배의 발효숙성 촉진에 관한 종합적 연구)

  • Bae, H.W.
    • Applied Biological Chemistry
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    • v.13 no.1
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    • pp.1-27
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    • 1970
  • The process of the forced aging of flue-cured tobacco leaves were studied extensively from various scientific points of view. The Flue-cured tobacco leaves were inoculated and fermented with nicotine resistant Hansenula yeast, or the leaves were subjected under simple forced aging. The above two processes of forced aging were studied from the summarized points of microbiology, physics, chemistry, and biochemistry, and the resulted products ware compared in their physical, chemical and biochemical quality determining factors with that of raw material tobacco leaves (dried-tobacco leaves) and 2 years aged high quality tobacco leaves. The summary results were as follows. 1) The Korean flue-cured tobacco leaves, were forcedly aged under the basic optimum aging condition, temperature $40^{\circ}C$, moisture contents 18%, relative humidity 74%. It was found that this aging condition was the best in bringing the quality of forcedly aged tobacco leaves to the utmost state. 2) Under this optimum temperature and moisture condition of forced aging in about 20 days the forcedly aged tobacco leaves both with yeast inoculation and without yeast inoculation showed the equivalent tobacco qualities comparable with that of more than 2 years aged tobacco leaves. 3) The forcedly aged tobacco leaves both with and without yeast inoculation under $40^{\circ}C$ temperature and $74^{\circ}C$ relative humidity achieved the necessary quality determining physical and chemical changes in about 20 days. 4) The microbial changes during the forced aging were as follows. The population of yeasts and bacteria increased until to 15 days of aging, then decreased thereafter. Whereas the molds grew continously until the end of fermentation. 5) The tobacco quality determing physico-chemico-properties of yeast inoculated aged and simple forcedly aged tobacco leaves, progressed as the follows in time. As the forced aging progresses, swelling and combustibility properties were improved. The pH, total reducing materials, total sugars, alkaloids contents decreased. The contents of organic and ether extractable materials increased. The total nitrogen, protein, crude fiber, ash contents showed no changes. The color properties, excitation purity, luminance, main wave length, showed equivalent changes comparable with that of 2 years aged tobacco leaves. 6) The changes in chemical components in yeast treated and simple forcedly aged tobacco leaves during $15{\sim}20{\;}days$ of forced aging were as follows. The following chemical components decreased as the aging. Sugars-sucrose. rhamnose, glucose. Pigments-chlorophyll, carotenes, xanthophyll and violax anthine. Polyphenols-rutin, chlorogenic and, coffeic acid. Organic acids-iso-butylic, crotonic, caprylic, galacturonic, tartaric, succinic, citric acid. Alkaloids-nicotine, nornicotine. The following components increased as the forced aging progressed. Sugars-frutose, maltose, raffinose. Amino acids-proline, cystine. Organic acids-formic, acetic, propionic, n-butyric, iso-valeric, n-valeric, malic, oxalic, malonic, ${\alpha}-ketoglutaric$, fumaric, glutaric acid. 7) During the forced aging of tobacco Leaves the oxygen-uptake decreased gradually. The enzyme activities of polyphenol oxidase, ${\beta}-amylase$ ${\alpha}-amylase$ decreased gradually. The activities of the enzymes, catalase, and invertase increased once then decreased at the later stage.

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