• Title/Summary/Keyword: Continuous cultures

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Semi-continuous cultivation of the mixotrophic dinoflagellate Gymnodinium smaydae, a new promising microalga for omega-3 production

  • Lim, An Suk;Jeong, Hae Jin;You, Ji Hyun;Park, Sang Ah
    • ALGAE
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    • v.35 no.3
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    • pp.277-292
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    • 2020
  • Omega-3 fatty acids, including eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), are polyunsaturated fatty acids beneficial to human health. A limited number of microalgae have been used for commercial omega-3 production, which necessitates the identification of new microalgae with high omega-3 contents. We explored the fatty acid composition and EPA and DHA contents of the mixotrophic dinoflagellate Gymnodinium smaydae fed with the optimal algal prey species Heterocapsa rotundata. Cells of G. smaydae were found to be rich in omega-3 fatty acids. In particular, the DHA content of G. smaydae was 21 mg g-1 dry weight, accounting for 43% of the total fatty acid content. The percentage of DHA in the total fatty acid content of G. smaydae was the highest among the reported microalgae except for Crypthecodinium cohnii. Moreover, to determine if the prey supply interval affected the growth rate of G. smaydae and its fatty acid content, three different prey supply intervals (daily, once every 2 d, and once for 4 d) were tested. Daily prey supply yielded the highest total fatty acid and DHA contents in G. smaydae. Furthermore, we successfully produced high-density G. smaydae cultures semi-continuously for 43 d with daily prey supply. During the semi-continuous cultivation period, the highest density of G. smaydae was 57,000 cells mL-1, with an average growth rate of 0.7 d-1. Taken together, the percentage of EPA and DHA in the total fatty acid content was maintained in the range of 54.2-56.9%. The results of this study support G. smaydae as a promising microalgal candidate for commercial DHA production and demonstrate that daily supply of prey can efficiently produce high-density G. smaydae cultures for more than a month.

Improved Production of Live Cells of Lactobacillus rhamnosus by Continuous Cultivation using Glucose-yeast Extract Medium

  • Ling Liew Siew;Mohamad Rosfarizan;Rahim Raha Abdul;Wan Ho Yin;Ariff Arbakariya Bin
    • Journal of Microbiology
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    • v.44 no.4
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    • pp.439-446
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    • 2006
  • In this study, the growth kinetics of Lactobacillus rhamnosus and lactic acid production in continuous culture were assessed at a range of dilution rates $(0.05 h^{-1}\;to\;0.40h^{-1})$ using a 2L stirred tank fermenter with a working volume of 600ml. Unstructured models, predicated on the Monod and Luedeking-Piret equations, were employed to simulate the growth of the bacterium, glucose consumption, and lactic acid production at different dilution rates in continuous cultures. The maximum specific growth rate of L. rhamnosus, ${\mu}_{max}$, was estimated at $0.40h^{-1}$I, and the Monod cell growth saturation constant, Ks, at approximately 0.25g/L. Maximum cell viability $(1.3{\times}10^{10}CFU/ml)$ was achieved in the dilution rate range of $D=0.28h^{-1}\;to\;0.35h^{-1}$. Both maximum viable cell yield and productivity were achieved at $D=0.35h^{-1}$. The continuous cultivation of L. rhamnosus at $D=0.35h^{-1}$ resulted in substantial improvements in cell productivity, of 267% (viable cell count) that achieved via batch cultivation.

Cell Recycled Culture of Succinic Acid-Producing Anaerobiospirillum succiniciproducens Using an Internal Membrane Filtration System

  • Lee, Pyung-Cheon;Lee, Sang-Yup;Chang, Ho-Nam
    • Journal of Microbiology and Biotechnology
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    • v.18 no.7
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    • pp.1252-1256
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    • 2008
  • Cell recycled culture of succinic acid-producing Anaerobiospirillum succiniciproducens was anaerobically carried out using an internal membrane filter module in order to examine the physiological response of A. succiniciproducens to a high-cell-density environment. The optimal growth of A. succiniciproducens and its enhanced succinic acid productivity were observed under $CO_2$-rich conditions, established by adding $NaHCO_3$ and $Na_2CO_3$, in the cell recycled system. A. succiniciproducens grew up to 6.50 g-DCW/l, the highest cell concentration obtained so far, in cell recycled cultures. The cells did not change their morphology, which is known to be easily changed in unfavorable or stress environments. The maximum productivity of succinic acid was about 3.3 g/l/h, which is 3.3 times higher than those obtained in batch cultures. These results can serve as a guide for designing highly efficient cell recycled systems for succinic acid at a commercial level.

A Study on Industrial Media for Production of Lactic acid in Batch and Continuous Fermentations (회분식 및 연속배양에 있어서 고농도 젖산의 생산을 위한 공업용 배지연구)

  • 김양훈;이기범;문승현
    • KSBB Journal
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    • v.14 no.2
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    • pp.181-187
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    • 1999
  • We have investigated industrial media for lactic acid fermentation to reduce the cost of nitrogen sources. Corn steep liquor (CSL) was successfully used at 5% (v/v) in batch fermentations. Use of soluble CSL improved the productivity about 20% with an advantage of clearer fermentation broth. Yeast extract-complemented CSL improved the productivity about 20% with an advantage of clearer fermentation broth. Yeast extract-complemented CSL media further increased the increased the productivity. It was found that 3.1 g/L yeast extract and 5% CSL could be an effective substitute for 15 g/L yeast extract in 10% glucose medium. Brewing yeast was also used as a sole nitrogen source equivalent to 5% CSL. A continuous culture coupled with cell-recycle by microfiltration at the dilution rate of 0.05-0.065 h-1 led to the highest lactic acid productivity. Lactic acid was recovered by electrodialysis from the cell free broth. Depleted cell free broth supplemented with 5-10 g/L of yeast extract performed reasonably in batch and continuous cultures. Reuse of the fermentation broth may reduce the cost of raw materials as well as minimize the fermentation wastes.

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Continuous Production of Natural Colorant, Betacyanin, by Beta vulgaris L. Hairy Root

  • Kim, Sun-Hee;Ahn, Sang-Wook;Bai, Dong-Kyu;Kim, Kwang-Soo;Hwang, Baik;Lee, Hyeon-Yong
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.716-721
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    • 1999
  • It has been known that continuous cultivation of hairy root is difficult to maintain for a long period of time compared to the microbial and callus cultures. Chemostat cultivation was successfully carried out in order to economically produce a plant-based colorant, betacyanin, from red beet hairy root for more than 85 days in a 14-1 fermentor. The result from the chemostat cultivation was compared to those of the batch and fed-batch cultivations of red beet hairy roots. It was shown that hairy root reached its steady state within 50 days of the cultivation, and then maintained for about 25-30 days in a wide range of dilution rates. Total betacyanin production from the continuous process was also calculated to be 2.65g at 0.28(l/d) of dilution rate, compared to 0.196g from fed-batch cultivation. It was found that betacyanin production was a partially growth related process, yielding 0.376 mg/g-fresh wt. cell and $1.89{\times}10^{-5}$ mg/g-fresh wt. cell/d, with 0.92 of correlation factor in a partial growth-product model. It was also shown that the cell growth required was relatively large for maintenance amount of energy at a low dilution rate. The growth of hairy root was inhibited by high light intensity in following a photo-inhibition model. The growth parameters were estimated to be 0.3(l/d), $10.56kcal/\textrm{m}^2/h$,{\;}and{\;}35.81kcal/\textrm{m}^2/h$ for the maximum specific growth rate, half saturation light intensity, and inhibition light intensity, respectively.

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재조합 효모를 이용한 항혈전 단백질 히루딘 발효 생산공정의 최적화

  • Kim, Myeong-Dong;Gang, Hyeon-A;Lee, Sang-Gi;Seo, Jin-Ho
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.99-102
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    • 2001
  • Recombinant Saccharomyces cerevisiae strains harboring various copy numbers of hirudin gene were developed to study dependency of hirudin expression level on its gene copy number. A linear relationship between the copy number of hirudin expression cassette and hirudin expression level was observed up to 10 copies. A double <5-integration vector truncated wi 디 1 the unnecessary bacterial genes before yeast transformation showed a four-fold increase in transformation efficiency and a 1.3-fold enhancement in hirudin expression level compared with a single <5 system. Gratuitous hirudin expression strain was developed by disrupting the GALl gene of S. cerevisiae. Glucose that was fed in a limited manner effectively supported cell growth and hi겨din expression by the gratuitous strain. Effects of methanol concentrations on hirudin production in recombinant Hansenula polymorpha were investigated in continuous and fed-batch cultures. At a steady-state of continuous culture, an optimum methanol concentration of 1.7 g/L was determined at a dilution rate of 0.18 $h^{-1}$ with 1.8 mg/L ${\cdot}$ h hirudin productivity.

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Controlling Mammalian Cell Metabolism in Bioreactors

  • Hu, Wei-Shou;Weichang, Zhou;Lilith F. Europa
    • Journal of Microbiology and Biotechnology
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    • v.8 no.1
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    • pp.8-13
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    • 1998
  • Animal cells in culture typically convert most of the glucose they consume into lactate. The accumulation of lactate, however, is commonly cited as one of the factors that inhibit cell growth and limit the maximum cell concentration that can be achieved in culture. The specific production of lactate and the amount of glucose converted to lactate can be reduced when cells are grown in a fed-batch culture in which the residual glucose concentration is maintained at low levels. Such a fed-batch culture was used to grow and adapt hybridoma cells into a low-lactate-producing state before changing into continuous culture. The cells reached and maintained a high viable cell concentration at steady state. In a similar manner, cells that were initially grown in batch culture and a glucose-rich environment reached a steady state with a cell concentration that is much lower. The feed composition and dilution rates for both cultures were similar, suggesting steady state multiplicity. From a processing perspective the desired steady state among those is the one with the least metabolite production. At such seady state nutrient concentration in the feed can be further increased to increase cell and product concentrations without causing the metabolite inhibitory effect typically seen in a cell culture. Controlling cell metabolism in a continuous culture to reduce or eliminate waste metabolite production may significantly improve the productivity of mammalian cell culture processes.

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미세조류의 Methane 발효특성

  • 강창민;최명락
    • Microbiology and Biotechnology Letters
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    • v.24 no.5
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    • pp.597-603
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    • 1996
  • This study was carried out to examine degradation characteristics of microalgae Chlorella vulgaris in methane fermentation. We measured COD and VS reduction, gas and methane productivity, VFA (volatile fatty acid), respectively. Then we calculated material balance and hydrolysis rates in soluble and solid material. The substrate concentration was controlled from 14 gCOD$_{cr}$/l to 64 gCOD$_{cr}$/l in batch cultures, and HRT (hydraulic retention time) controlled from 2 days to 30 days in continuous experi- ments. The results were as follows. In batch culture, accumulated gas productivity increased with the increase of the substrate concentration. The SS and VSS was removed all about 30% increase of substrate concentration and the most of the degradable material removed during the first 10 days. The curve of gas and methane production rate straightly increased until substrate concentration is 26 gCOD$_{cr}$/l. In continuous culture experiments, the removal rates at HRT 10days were 20% for total COD and TOC, respectively. At longer HRT, there was no increase in the removal efficiency. At HRT 15 days, the removal rates were 30% for SS and VSS, respectively. Soluble organic materials were rapidly degraded, and so there was no accumulated. Soluble COD concentration was not increase regardless of HRT-increasing. That meaned the hydrolysis was one of the rate-limiting stage of methane fermentation. The first-order rate constants of hydrolysis were 0.23-0.28 day$^{-1}$ for VSS, and 0.07-0.08 day$^{-1}$ for COD.

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Reduction of Hexavalent Chromium by Escherichia coli ATCC 33456 in Batch and Continuous Cultures

  • Bae, Woo-Chul;Kang, Tae-Gu;Kang, In-Kyong;Won, You-Jung;Jeong, Byeong-Chul
    • Journal of Microbiology
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    • v.38 no.1
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    • pp.36-39
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    • 2000
  • Toxic hexavalent chromium, Cr(VI), was reduced to a less toxic trivalent chromium form by E. coli ATCC 33456. The suitable electron donor for Cr(VI) reduction was glucose. E. coli ATCC 33456 was more resistant to metal cations than other reported Cr(VI) reducing microorganisms. Cell growth was inhibited by the presence of Cr(VI) in a liquid medium and Cr(VI) reduction accompanied cell growth. With a hydraulic retention time of 20 h, Cr(VI) reducing efficiency was 100% to 84% when Cr(VI) concentration in the influent was in the range of 10 to 40 mg L$\^$-1/. Specific rate of Cr(VI) reduction was 2.41 mg Cr(VI) g DCW$\^$-1/ h$\^$-1/ when 40 mg L$\^$-1/ of Cr(VI) influent was used. This result suggested the potential application of E. coli ATCC 33456 for the detoxification of Cr(VI) in Cr(VI) contaminated wastewater.

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Effect of Partial Oxygen Pressure on the Growth and Defense Enzyme Activities of Streptomyces coelicolor in continuous culture system (Streptomyces coelicolor의 연속 배양시 산소 분압에 따른 방어 효소의 활성 변화)

  • 박용두;이계준;노정혜
    • Microbiology and Biotechnology Letters
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    • v.22 no.5
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    • pp.538-543
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    • 1994
  • Effect of partial oxygen pressure on the cell growth and the activities of oxidative defense enzymes were measured in the continuous culture of Streptomyces coelicolor. Both the wild type and the mutant strain resistant to hydrogen peroxide were cultured and the dry cell weight of the two cultures were measured at different oxygen tensions. Growth of the wild type was inhibited by oxygen at above 0.5 vvm. Growth of the hydrogen peroxide resistant mutant was stimulated by pure oxygen at 0.5 vvm but was inhibited by oxygen at 1.0 vvm. Therefore, growth of the hydrogen peroxide resistant mutant was less affected by the deleterious oxidative stress of oxygen. Activities of the several defense enzymes were also measured at different oxygen tensions. Activities of catalase and glucose-6-phosphate dehydrogenase increased significantly as oxygen pressure increased in the wild type culture. In the mutant, however, increase in those enzyme activities was not obvious whereas the uninduced levels of the above enzymes were higher than those of wild type. As judged by Western blotting, the amount of the major catalase increased as the oxygen pressure increased. This indicates that the induction of the catalase activity by oxygen pressure is mostly due to the increase in the expression level for the major catalase.

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