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Single Cell Oil Production from Undetoxified Arundo donax L. hydrolysate by Cutaneotrichosporon curvatus

  • Di Fidio, Nicola;Liuzzi, Federico;Mastrolitti, Silvio;Albergo, Roberto;De Bari, Isabella
    • Journal of Microbiology and Biotechnology
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    • v.29 no.2
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    • pp.256-267
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    • 2019
  • The use of low-cost substrates represents one key issue to make single cell oil production sustainable. Among low-input crops, Arundo donax L. is a perennial herbaceous rhizomatous grass containing both C5 and C6 carbohydrates. The scope of the present work was to investigate and optimize the production of lipids by the oleaginous yeast Cutaneotrichosporon curvatus from undetoxified lignocellulosic hydrolysates of steam-pretreated A. donax. The growth of C. curvatus was first optimized in synthetic media, similar in terms of sugar concentration to hydrolysates, by applying the response surface methodology (RSM) analysis. Then the bioconversion of undetoxified hydrolysates was investigated. A fed-batch process for the fermentation of A. donax hydrolysates was finally implemented in a 2-L bioreactor. Under optimized conditions, the total lipid content was 64% of the dry cell weight and the lipid yield was 63% of the theoretical. The fatty acid profile of C. curvatus triglycerides contained 27% palmitic acid, 33% oleic acid and 32% linoleic acid. These results proved the potential of lipid production from A. donax, which is particularly important for their consideration as substitutes for vegetable oils in many applications such as biodiesel or bioplastics.

Nitroglycerin Biodegradation under Denitrification Conditions and Corresponding Microbial Community Shifts upon Acclimation (탈질조건에서 nitroglycerin의 생물학적 분해 동역학 및 미생물 군집 변화)

  • Choi, Wonchul;Bae, Bumhan
    • Journal of Soil and Groundwater Environment
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    • v.24 no.5
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    • pp.42-54
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    • 2019
  • Biodegradation of an explosive compound, glyceryl trinitrate (GTN), was studied with a denitrifying microbial culture grown in a sequencing batch reactor and a GTN acclimated denitrifying culture. The GTN acclimated culture, which were fed on GTN for 1 month, degraded GTN regioselectively via denitration on C1 position as compared to C2 position denitration by denitrifying culture that has never been exposed to GTN. Accumulation of two isomeric glyceryl dinitrates (GDNs) in both culture medium suggests that GDN denitration is the rate-limiting step in GTN biodegradation. The first order GTN degradation rate normalized to cell concentration of the acclimated culture was calculated to be 0.045 (${\pm}0.002$) L/g-hr. Increasing concentration of electron acceptor(nitrate) resulted in discouraged GTN degradation. According to microbial community analysis, prolonged GTN exposure resulted in 25% increase in the genus level of the GTN acclimated culture with the disappearance of two dominating denitrifying microbial species of Methyloversatilis universalis and Hyphomicrobium zavarzinii in the denitrifying culture.

The Overall Performance Improvement of Microbial Fuel Cells Connected in Series with Dairy Wastewater Treatment

  • Choudhury, Payel;Bhunia, Biswanath;Bandyopadhyay, Tarun Kanti;Ray, Rup Narayan
    • Journal of Electrochemical Science and Technology
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    • v.12 no.1
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    • pp.101-111
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    • 2021
  • To improve the potential of single chamber microbial fuel cells (SCMFCs) as an applicable technology, the main challenge is a practical application for larger scales bioenergy production from potent exoelectrogenic microorganism with real dairy wastewater. To increase power generation, three individual MFCs were together operated in series best under the fed batch condition for 15 days. The volume of MFC 1 and MFC 2 is "300 mL" and MFC 3 is "500 mL" respectively. The individual MFCs 1, MFC 2 and MFC 3 gives an open circuit voltage of 0.60 V, 0.66 V and 0.55 V and result in total working voltage when connected in series of 1.745V, which lead an LED to glow. The maximum power densities obtained from MFC 1, MFC 2 and MFC 3 are 62 mW/㎡, 50 mW/㎡ and 45 mW/㎡ (normalized to the surface area of the anodic electrode, which was 50 ㎠ for all three MFCs), and corresponding to current densities of 141 mA/㎡, 155 mA/㎡ and 123 mA/㎡, respectively. Therefore this work suggests the cheapest way to connect microbial fuel cells in series to gain power with the lowest operating cost and chemical oxygen demand (COD) removal.

Cultivation Condition of Transformant Alcaligenes eutrophus Harboring Cloned phbC Gene for Production of P(3-hydroxybutyrate-3-hydroxyvalernte) Containing High Molar Fraction of 3-Hydroxyvalerate. (P(3-hydroxybutyrate-3-hydroxyvalerate)의 생산을 위한 재조합 phbC 유전자를 형질전환시킨 Alcaligenes eutrophus의 배양조건 검토)

  • 권순일;정영미;이용현
    • Microbiology and Biotechnology Letters
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    • v.26 no.6
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    • pp.537-544
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    • 1998
  • The cultivation conditions of transformant Alcaligenes eutrophus AER5 harboring cloned phbC gene for mass production of poly (3-hydroxybutyrate-3-hydroxyvalerate)[P(3HB-3HV)] containing high molar fraction of 3-hydroxyvalerate (3-HV) were investigated. In two-stage batch cultivation, transformant accumulated P(3HB-3HV) containing 52.2 mol% of 3HV compared to 30 mol% of parent strain A. eutrophus H16. The increased 3-HV molar fraction was due to the amplified activity of PHB synthase participating in condensation of 3-HB and 3-HV. To increase efficiency of P(3HB-3HV) accumulation, fructose was added along with precursor compound valerate, and total cell mass and P(3HB-3HV) concentrations remarkably increased, but not 3-HV molar fraction. The effect of magnesium ion showed that P(3HB-3HV) concentration and 3-HV molar fraction were significantly increased upto 6.1 g/L and 71.3 mol% at 0.01 g/L of MgSO$_4$, respectively. The efficiency of several pH adjuster, NaOH, NaOH and (NH$_4$)$_2$SO$_4$, and NH$_4$OH, on total cell mass, p(3HB-3HV) concentration, and 3-HV molar fraction was also compared. To overcome the disadvantage of two-stage cultivation, one-stage intermittent fed-batch cultivation was attempted, such a way 10.0 g/L of fructose was supplied for cell growth at initial 36 hr and then 10.0 g/L of valerate and 5.0 g/L of fructose were applied to induce the accumulation of P(3HB-3HV), consequently, 10.4 g/L of P(3HB-3HV) with 38 mol% of 3-HV fraction could be obtained after 72 hr. These results can be used for elucidating cultivation strategy for mass production of P(3HB-3HV) containing high 3-HV molar fraction using transformant A. eutrophus AER5 harboring cloned phbC gene.

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Evaluation of biological treatment of cutting-oil wastes using sequencing batch reactor (SBR) process (연속 회분식 반응조 (SBR) 공정을 이용한 폐절삭유의 생물학적 처리능 평가)

  • Baek, Byung-Do;Kim, Chang-Seop;Kim, Jun-Young;Chang, In-Soung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.10 no.7
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    • pp.1654-1660
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    • 2009
  • Two different cutting-oils from H and B companies which are sold as an eco-friendly cutting-oils were selected and the biodegradability of these commercially available cutting-oils was evaluated by the sequencing batch reactor (SBR) processes. The cutting-oil wastes ($H_1$) pre-treated by coagulation/flocculation was used as an influent to SBR. When the F/M ratio was operated 0.04 to 0.08kgCOD/kgMLSS d, removals of $BOD_5$and $COD_{Cr}$were above 97% and 91%, respectively. T-N and T-P removals were above 76% and 81%, respectively. If the diluted cutting-oil wastes ($B_1$) was used as an influent of the SBR, $COD_{Cr}$removals were above 77% at the F/M ratio of 0.01-0.02kgCOD/kgMLSS d. After the cutting-oil wastes was treated by coagulation/ flocculation ($B_2$), $COD_{Cr}$removals was above 85%. If the pre-treated cutting-oil wastes were mixed with a synthetic wastewater ($B_3$) and fed into the SBR in order to mimic the real wastewater treatment plant situation, $BOD_5$and $COD_{Cr}$removals were above 97%, 91%, respectively. T-N and T-P removals were above 79% and 76%. The ratio between $BOD_5$and $COD_{Cr}$, ($COD_{Cr}$-$BOD_5$)/$COD_{Cr}$, indicating the biodegradability of effluent of the SBR, was calculated to 85% and 61%. This means that significant amounts of non-readily-biodegradable organic compounds in the effluent of $H_1$, $B_3$are still present.

Microbial Communities of Activated Sludge in an Anaerobic/Aerobic Sequencing Batch Reactor using Slot Hybridization (Slot Hybridization을 이용한 연속 회분식 반응기내 미생물 분포 조사)

  • Jeon, Che Ok;Shin, Kum-Joo;Lee, Dae Sung;Suh, Pann-Ghill;Park, Jong Moon
    • Journal of Korean Society of Environmental Engineers
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    • v.22 no.5
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    • pp.939-947
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    • 2000
  • Enhanced biological phosphorus removal (EBPR) was performed in an anaerobic/aerobic sequencing batch reactor (SBR). Influent was a synthetic wastewater based on acetate as a carbon source. The sludge age and hydraulic retention time were kept at 10 days and 16 hrs, respectively, Phosphate release during the anaerobic period and phosphate uptake in aerobic period were increased gradually with time. and after about 200 days, steady-state operation could be achieved with complete removal of influent phosphate. Number distribution of microbial community in the sludge performing EBPR was investigated during the steady state operation. 17 rRNA targeted oligonucleotide probes were designed and slot hybridization technique was used to determine the number distribution of each microorganism. In the acetate fed SBR, rRNA belonging to the beta subclass of proteobacteria was the most dominant in total rRNA and rRNA matching to CTE probe was the second, rRNAs of Acinetobacter, Aeromonas and Pseudomonas, which are usually thought as phosphorus accumulating organisms in EBPR processes, constituted less than 10% of total rRNA. From this community analysis, it was inferred that microorganisms belong to the beta subclass of proteobacteia (BET) and CTE such as Rhodocyclus group were important in biological phosphorus removal. Therefore, the role of Acinetobacter, Aeromonas and Pseudomonas in the EBPR might have been overestimated.

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Production of L-Lactic Acid from Soluble Starch by Enterococcus sp. JA-27. (Enterococcus sp. JA-27에 의한 가용성 전분으로부터 L형 젖산의 생산)

  • 김경아;김미경;장경린;전홍기
    • Microbiology and Biotechnology Letters
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    • v.31 no.3
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    • pp.250-256
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    • 2003
  • Lactic acid bacteria with amylolytic and acid producing activities can ferment starch directly to lactic acid thereby producing a monomer for the production of biodegradable poly lactic acid (PLA). In this study, the strain producing L-lactic acid from soluble starch was isolated from Nuruk. The isolated strain was identified as Enterococcus sp. through its morphological, cultural, biochemical characteristics as well as the 16S rDNA sequence analysis, and named Enterococcus sp. JA-27. Enterococcus sp. JA-27 produced exclusively L-lactic acid from soluble starch as a carbon source. The optimal conditions for the maximum production of L-lactic acid from Enterococcus sp. JA-27 were 30 C, pH 8, 1.5 % soluble starch as a substrate and 3.5 % tryptone as a nitrogen source, 0.1 % $K_2$$HPO_4$, 0.04 % $MgSO_4$. $7H_2$O, 0.014 % $MnSO_4$$.$4$H_2O$, 0.004% $FeSO_4$$.$$7H_2$O. Batch and fed batch culture were carried out and the former was more effective. L-Lactic acid production in the optimum medium was significantly increased in a 7 L jar fermenter, where the maximum L-lactic acid concentration was 3 g/L. For the purification of lactic acid in fermented broth, two stage ionexchange column chromatographies were employed and finally identified by HPLC.

Starter culture production of Rhodospirillum rubrum P17 for use in treatment of organic waste water (유기폐수처리를 위한 Rhodospirillum rubrum P17의 종균생산)

  • Cho, Kyung-Dug;Kang, Seong-Og;Lim, Wang-Jin;Cho, Hong-Yon;Yang, Han-Chul
    • Applied Biological Chemistry
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    • v.36 no.6
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    • pp.488-494
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    • 1993
  • A photosynthetic bacterium strain P17 having high growth rate and assimilating ability of organic acids was isolated from several soil samples, which was identified as Rhodospirillum rubrum. Cultural conditions of the strain P17 were examined for the production of starter culture used in the treatment of organic waste water. The addition of organic acids mixture as carbon source containing 0.2% Na-acetate, 0.1% Na-propionate and 0.2% Na-lactate and 0.1% of yeast extract as growth factor stimulated the cell growth. The maximal cell production was obtained at $30^{\circ}C$, pH 7.0, 2,500 lux of illumination and $50{\sim}100\;rpm$ of agitation. Under the optimal conditions of batch and fed-batch culture systems in a Jar fermentor, 5.17 g/l and 7.93 g/l of cells were obtained after S days of cultivation, respectively. In continuous culture system, the cell productivity was 0.206 g/l/h at a dilution rate of 0.21 $h^{-1}$. When R. rubrum P17 was cultivated in a soybean curd waste water, initial COD level(3,240 mg/l) of the waste water was reduced to 250 mg/l after 4 days of cultivation.

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Accumulation of Poly-$\beta$-Hydroxybutyrie Acid by Alcaligenes sp. (Alcaligenes sp.에 의한 Poly-$\beta$-Hydroxybutyric Acid의 축적)

  • 임명순;손홍주;박수민;이종근;이상준
    • Microbiology and Biotechnology Letters
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    • v.20 no.4
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    • pp.363-370
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    • 1992
  • Microorganisms capable of accumulating poly-p-hydroxybutyric acid(PHB) were isolated from soil by enrichment culture technique. Among them, the strain designated as FL-027 had high PHB productivity and was identified as Alcaligenes. The optimal medium composition for cell growth was 8.0 $g/\ell$ of fructose and 3.0 $g/\ell$ of $(NH_4)_2S0_4$, equivalent to C/N ratio 5.04 at pH 7.0 and $30^{\circ}C$. To investigate the optimal conditions for the PHB accumulation, we divided the process into two stages; the first stage for the growth of the cell in nutrient-rich medium and the second stage for the PHB accumulation in nutrient-deficiency medium. The optimal conditions for PHB accumulation were 8.0 $g/\ell$ of fructose and 0.25 $g/\ell$ of $(NH_4)_2S0_4$, equivalent to C/N ratio 60 at pH 6.5 and $30^{\circ}C$. PHB accumulation was stimulated by deficiency of nutrients such as $NH_4^+$, $Ca^{2+}$, $SO_4^{2+}$ in medium. Among them. $NH_4^+$ deficiency was chosen because of its effectiveness. We found the inhibition of cell growth by fructose in batch culture. In order to keep the fructose concentration at an optimal level, intermittent feeding fed-batch culture was employed, and the cell concentration as high as 10.83 $g/\ell$ whose PHB content was responsible for 43% of the dry cell weight. The purified PHB was identified as homopolymer of 3-hydroxybutyric acid by using IR and $^1H-NMR$.

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Supplementing Rhodobacter sphaeroides in the diet of lactating Holstein cows may naturally produce coenzyme Q10-enriched milk

  • Bae, Gui-Seck;Choi, Ahreum;Yeo, Joon Mo;Kim, Jong Nam;Song, Jaeyong;Kim, Eun Joong;Chang, Moon Baek
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.1
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    • pp.40-46
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    • 2018
  • Objective: To examine the effects of Rhodobacter sphaeroides (R. sphaeroides) supplementation as a direct-fed microbial (DFM) on rumen fermentation in dairy cows and on coenzyme Q10 (CoQ10) transition into milk, an in vitro rumen simulation batch culture and an in vivo dairy cow experiment were conducted. Methods: The characteristics of in vitro ruminal fermentation were investigated using rumen fluids from six cannulated Holstein dairy cows at 2 h post-afternoon feeding. A control treatment was included in the experiments based on a typified total mixed ration (TMR) for lactating dairy cows, which was identical to the one used in the in vivo study, plus R. sphaeroides at 0.1%, 0.3%, and 0.5% TMR dry matter. The in vivo study employed six ruminally cannulated lactating Holstein cows randomly allotted to either the control TMR (C-TMR) treatment or to a diet supplemented with a 0.5% R. sphaeroides culture (S-TMR, dry matter basis) ad libitum. The presence of R. sphaeroides was verified using denaturing gradient gel electrophoresis (DGGE) applied to the bacterial samples obtained from the in vivo study. The concentration of CoQ10 in milk and in the supernatant from the in vitro study was determined using high performance liquid chromatography. Results: The results of the in vitro batch culture and DGGE showed that the concentration of CoQ10 significantly increased after 2 h of R. sphaeroides supplementation above 0.1%. When supplemented to the diet of lactating cows at the level of 0.5%, R. sphaeroides did not present any adverse effect on dry matter intake and milk yield. However, the concentration of CoQ10 in milk dramatically increased, with treated cows producing 70.9% more CoQ10 than control cows. Conclusion: The CoQ10 concentration in milk increased via the use of a novel DFM, and R. sphaeroides might be used for producing value-added milk and dairy products in the future.