• Title/Summary/Keyword: Two-phase fermentation

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Evaluation of Extractants for Bio-butanol Extraction Fermentation Using Organic Solvents and Ionic Liquids (유기용매와 이온성액체를 이용한 바이오 부탄올 추출발효 용매 선정 평가)

  • Cho, Min-Ok;Lee, Sun-Mi;Sang, Byoung-In;Um, Young-Soon
    • KSBB Journal
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    • v.24 no.5
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    • pp.446-452
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    • 2009
  • Oleyl alcohol, butyl butyrate, and two different ionic liquids were evaluated for the extraction of butanol from culture broth without toxic effect to cells. The tested solvents showed more than 50% extraction efficiency, and oleyl alcohol was chosen as the best extractant for butanol among the used extractants with a partition coefficient of 2.89. When oleyl alcohol was used as an extractant, more than 80% of butanol was extracted in the wide range of butanol concentrations (1-20 g/L) and pH values (pH 4-5.5). In extractive fermentation using oleyl alcohol only, there was 11% more butanol production and glucose consumption when compared to that without extractive fermentation, implicating a reduced inhibitory effect of butanol due to butanol removal to the oleyl alcohol phase. In addition, oleyl alcohol did not inhibit cell growth, while a mixture of oleyl alcohol and butyl butyrate with the volume ratio of 9:1~7:3 inhibited either butanol production or cell growth significantly due to the toxicity of butyl butyrate to cells. In conclusion, oleyl alcohol can be used as an efficient and non-toxic solvent for extractive fermentation for butanol production.

Mathematical Modeling with Cell Morphology and Its Application to Fed-batch Culture in Cephalosporium Fermentation (Cephalosporium 발효시 균체의 형태학적 측면을 고려한 수학적 모델링 및 유가식 배양에의 응용)

  • 김의용;유영제
    • Microbiology and Biotechnology Letters
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    • v.19 no.5
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    • pp.521-535
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    • 1991
  • A kinetic model incorporating cell morphology in cephalosporin C biosynthesis by Cephalosporium amemoniurn was developed. The double-substrate Double-substrate kinetic model was used to describe cell growth. Methionine controlled the rate of growth while glucose ultimately controlled the extent of growth. The changes in specific product formation rate were associated with morphologenesis, especially cell differentiation. To increase the productivity of cephalosporin C, the proposed model equations were applied to a fed-batch culture. The algorithm to optimize the fed-batch culture consists of two steps; cell growth was maximized in the growth phase and then cephalosporin C production was maximized in the production phase. The increase of about 33% in the cephalosporin C titre was obtained by the optimal feeding scheduling in comparison with that of batch culture.

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Fabrication and Characterisation of a Novel Pellicular Adsorbent Customised for the Effectvie Fluidised Bed Adsorption of Protein Products

  • Sun, Yam;Pacek, Andrzej W.;Nienow, Alvin W.;Lyddiatt, Andrew
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.6
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    • pp.419-425
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    • 2001
  • A dense pellicular solid matrix has been fabricated by coating 4% agarose gel on to dense zironia-silica(ZS) spheres by watr-in-oil emulsification . The agarose evenly laminated the ZS bead to a depth of 30㎛, and the resultin gpellicular assembly was characterised by densities up to 2.39g/mL and a mean particle dimeter of 136 ㎛. In comparative fluidisation tests, the pellicular solid phase exhibited a two-fold greater flow velocity than commercial benchmark ad-sorbents necessary to achieve common values of bed expansion. Furthermore, the perlicular parti-cles were characterised by improved qualities of chromatographic behaviour, particularly with re-spect to a three-fold increase in the apparent effective diffusivity of lysozyme within a pellicular assembly modified with Cibacron Blue 3GA. The properties of rapid protein adsorption/desorp-tion were attributed to the physical design and pellicular deployment of the reactive surface in the solid phase. When combined with enhanced feedstock throughput, such practical advantages recommend the pellicular assembly as a base matrix for the selective recovery of protein products from complex, particulate feedstocks(whole fermentation broths, cell disruptates and biological extracts).

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A Study on the Quality Characteristics of Kimchi with Mulberry Leaf Powder (뽕잎 분말을 첨가한 김치의 품질 특성에 관한 연구)

  • La, Sun-Hwa;Choi, Mi-Kyeong;Shin, Seung-Mee
    • The Korean Journal of Food And Nutrition
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    • v.20 no.1
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    • pp.53-62
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    • 2007
  • In this study, physicochemical, microbiological and organoleptic analyses were carried out on cabbage kimchi, a representative fermented food, that was made with 0.5%, 1.0%, and 1.5% mulberry leaf powder during fermentation. This kimchi was then compared to, kimchi without added mulberry leaf powder. The pH values showed minimal differences between the two types of kimchi at the beginning of fermentation. The total acidities were low in every plot of kimchi and increased according to the fermentation. The total microbial cell and Lactobacilus counts increased rapidly in the exponential phase according to the fermentation, and showed little increase in the stational phase. Among the analyzed plots, the lowest population was found in the kimchi containing 1.0% of mulberry leaf powder. This kimchi, in particular, also had the best quality scores, overall acceptance and organoleptic test results during fermentation. Fermentation was slowed in the kimchi with mulberry leaf powder, according to the amount of added as compared to the kimchi without it also showed less acidity. The color appearance, however, of the kimchi with added mulberry leaf powder, was inferior to that of the kimchi without mulberry leaf powder.

Production of Pyruvate Dehydrogenase Complex-E2 Specific Human Monoclonal Antibody in Fed-batch Culture Systems with High Cell Density Recombinant Escherichia coli (고농도 재조합 대장균의 Fed-batch 배양 시스템을 이용한 Pyruvate Dehydrogenase Complex-E2 특이성 인간 모노클론 항체의 생산)

  • 이미숙;전주미;차상훈;정연호
    • KSBB Journal
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    • v.15 no.5
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    • pp.489-496
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    • 2000
  • Several culture systems including batch, two-stage CSTR, semi-fed batch, and two-stage cyclic fed-batch were investigated for the efficient production of the Fab fraction of PDC-E2 specific human monoclonal antibody using high cell density recombinant E. coli. A two-phase batch system and a two-stage continuous system were examined to overcome plasmid instability problems, by separating the growth and the production stages. The cell density and productivity of the two-stage continuous culture was better than that of the two-phase batch fermentation. In the two-stage continuous culture system with DO-stat, the cell growth and the productivity were superior to those of the system without the DO control. Also, almost total plasmid stability was maintained in the two-stage continuous culture system. Modified M9 medium was selected as an optimum feeding medium for the fed-batch process, and the optimum C/N ratio determined to be 2:3. The optimum feeding rate was $0.6g/\ell/hr$ for a constant feeding strategy in semi-fed batch system. When the feeding medium was fed by pulsing, it was observed that more frequent pulsing resulted in improved cell growth. The linear feeding method was the most efficient of the various feeding methods tested. Finally, high cell density culture using a two-stage cyclic fed batch system with pH-stat was tried because the linear feeding method showed limitations in terms of obtaining high cell densities, and a cell density of $54 g/\ell$ was achieved. It was concluded that the two-stage cyclic fed batch system was the most efficient system for high cell density culture of the systems tested. However, productivity improvements were lower than expected due to the extremely high accumulations of acetate, although the low levels of residual glucose were maintained.

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폐지 슬러지를 이용한 혐기성 메탄발효 특성 분석

  • Jo, Geon-Hyeong;Kim, Jung-Gon;Jeong, Hyo-Gi;Kim, Seong-Jun;Kim, Si-Uk
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.367-370
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    • 2003
  • This study was carried out to investigate the possibility for reuse of solid organic wastes such as saccharified newspapers and boxes by two-phase anaerobic fermentation system. When 15g of newspaper and box wastes were digested for 24 days by batch fermentation, tCOD removal rate were found to be 60.9 and 62.4%, respectively. During this period, the amounts of biogas produced were 6.95 and 6.43L. The removal efficiencies of total solid were 34.8 and 33.4%, and those of volatile solid were 40.0 and 39.2%, respectively. That pH was around 7.5 after 20-days operation means methane fermentation is well advanced. In case of semicontinuous reaction, tCOD removal efficiencies of newspaper and box wastes were 64.7 and 65.0%, respectively for 14-days operation. It has been shown that each of the average biogas amounts produced after 25 days operation (stabilization stage for methane fermentation) was 0.31 and 0.30L/g dry wt./day, respectively, and each methane contents was 57.3 and 56.2%, respectively. After the reaction continued for 25 days, pHs in the anaerobic acidogenic and methanogenic fermenters were shown to be 5.0 and 7.5, respectively.

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Characteristics of Fibrinolytic Enzymes of Bacillus licheniformis CY-24 Isolated from Button Mushroom Compost (양송이 배지로부터 분리한 Bacillus licheniformis CY-24의 섬유소분해 효소의 특성)

  • Min, Gyeong-Jin;Park, Hea-sung;Lee, Een-ji;Lee, Chan-Jung
    • The Korean Journal of Mycology
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    • v.49 no.2
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    • pp.199-209
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    • 2021
  • The present study was performed to improve the technique used for fermenting the mushroom growth medium. Taxonomic analysis of 16S rDNA sequence from the predominant Bacillus strain CY-24 isolated during the fermentation phase of the rice straw medium identified it as Bacillus licheniformis. In addition, the growth environment of B. licheniformis was also examined in this study, which revealed the optimal growth temperature and pH to be 30 ℃ and 6.0, respectively. This study also revealed that carboxymethyl cellulase (CMCase) and polygalacturonase (PGase) enzymes isolated from B. licheniformis achieved their maximal activities at 50 ℃ and 60 ℃ respectively. Furthermore, the study confirmed that the two enzymes, i.e., CMCase and PGase in B. licheniformis are stable at temperatures above 60 ℃. The present study thus demonstrates that B. licheniformis CY-24 possesses excellent enzymatic properties. It also reveals that the action of enzymes during the production of growth mediums used for the cultivation of mushrooms is closely associated with the promotion of fermentation and softening of the rice straw. Overall, this study provides elementary information regarding the role of B. licheniformis enzymes during growth medium fermentation for Agaricus bisporus cultivation.

Comparison of Mcrobial and Physicochemical Properties between Pogi Kimchi and Mat Kimchi (포기김치와 맛김치의 미생물학적 및 이화학적 품질 특성 비교)

  • Moon, Eun Woo;Kim, Su-Yeon;Dang, Yun-Mi;Park, Boyeon;Park, Eun Jin;Song, Hye Yeon;Yang, Jisu;Yoon, So Ra;Seo, Hye-Young;Ha, Ji-Hyoung
    • Journal of the Korean Society of Food Culture
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    • v.34 no.2
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    • pp.217-223
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    • 2019
  • This study aimed to evaluate the characteristic differential between whole cabbage kimchi (pogi kimchi) and sliced cabbage kimchi (mat kimchi) during kimchi fermentation at $6^{\circ}C$. The difference of microbial and physicochemical properties was investigated until 6 weeks. For the changes in the microbial flora, both kimchi samples exhibited a continuous increase in total aerobic bacteria and lactic acid bacteria (LAB) population size up to 2 weeks followed by a stationary phase until 5 weeks. Interestingly, the number of LAB of mat kimchi was overall higher than that of pogi kimchi during kimchi fermentation. We speculate that mat kimchi has in a more advantageous growth condition than pogi kimchi for microbial growth because small kimchi cabbage size appropriately derives nutritional supply in order to increase the LAB growth. During lactic fermentation at $6^{\circ}C$, physicochemical changes in the pH, salinity, and titratable acidity was observed to be no significant differences between two types of kimchi. Furthermore the contents of organic acids such as oxalic acid, citric acid, malic acid, lactic acid, fumaric acid, and acetic acid was not significantly different (p>0.05) between both kimchi samples as well as the contents of total free amino acid.

Influence of Temperature and pH on Fermentation Pattern and Methane Production in the Rumen Simulating Fermenter (RUSITEC)

  • Bhatta, R.;Tajima, K.;Kurihara, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.3
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    • pp.376-380
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    • 2006
  • An experiment was conducted to study the effect of temperature and pH on in vitro nutrient degradability, volatile fatty acid profile and methane production. The fermenter used was the semi-continuous system, known as the rumen simulation technique (RUSITEC). Sixteen cylinders were used at one time with a volume of 800 ml, the dilution rate was set at 3.5%/hour, the infused buffer being McDougall's artificial saliva. Basal diet (9.6 g DM) used in RUSITEC consisted of (DM) 6.40 g Timothy hay, 1.86 g crushed corn and 1.34 g soybean meal. The food for the fermentation vessel was provided in nylon bags, which were gently agitated in the liquid phase. The experiment lasted for 17 d with all the samples taken during the last 5 d. Treatments were allocated at random to four vessels each and were (1) two temperature levels of $39^{\circ}C$ and $41^{\circ}C$ (2) two pH levels of 6.0 and 7.0. The total diet contained ($g\;kg^{-1}$ DM) 957 OM, 115 CP and $167MJ\;kg^{-1}$ (DM) GE. Although increase in temperature from $39^{\circ}C$ to $41^{\circ}C$ reduced degradation of major nutrients in vitro, it was non-significant. Interaction effect of temperature with pH also reflected a similar trend. However, pH showed a significant (p<0.05) negative effect on the degradability of all the nutrients in vitro. Altering the in vitro pH from 7 to 6 caused marked reduction in DMD from 60.2 to 41.8, CPD from 76.3 to 55.3 and GED from 55.3 to 35.1, respectively. Low pH (6) depressed total VFA production (61.9 vs. 34.9 mM) as well as acetate to propionate ratio in vitro (from 2.0 to 1.5) when compared to pH 7. Compared to pH 7, total gas production decreased from 1,841 ml to 1,148 ml at pH 6, $CO_2$ and $CH_4$ production also reduced from 639 to 260 ml and 138 to 45 ml, respectively. This study supported the premise that pH is one of the principal factors affecting the microbial production of volatile fatty acids and gas. Regulating the ruminal pH to increase bacterial activity may be one of the methods to optimize VFA production, reduce methane and, possibly, improve animal performance.

Recombinant Glargine Insulin Production Process Using Escherichia coli

  • Hwang, Hae-Gwang;Kim, Kwang-Jin;Lee, Se-Hoon;Kim, Chang-Kyu;Min, Cheol-Ki;Yun, Jung-Mi;Lee, Su Ui;Son, Young-Jin
    • Journal of Microbiology and Biotechnology
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    • v.26 no.10
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    • pp.1781-1789
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    • 2016
  • Glargine insulin is a long-acting insulin analog that helps blood glucose maintenance in patients with diabetes. We constructed the pPT-GI vector to express prepeptide glargine insulin when transformed into Escherichia coli JM109. The transformed E. coli cells were cultured by fed-batch fermentation. The final dry cell mass was 18 g/l. The prepeptide glargine insulin was 38.52% of the total protein. It was expressed as an inclusion body and then refolded to recover the biological activity. To convert the prepeptide into glargine insulin, citraconylation and trypsin cleavage were performed. Using citraconylation, the yield of enzymatic conversion for glargine insulin increased by 3.2-fold compared with that without citraconylation. After the enzyme reaction, active glargine insulin was purified by two types of chromatography (ion-exchange chromatography and reverse-phase chromatography). We obtained recombinant human glargine insulin at 98.11% purity and verified that it is equal to the standard of human glargine insulin, based on High-performance liquid chromatography analysis and Matrix-assisted laser desorption/ionization Time-of-Flight Mass Spectrometry. We thus established a production process for high-purity recombinant human glargine insulin and a method to block Arg (B31)-insulin formation. This established process for recombinant human glargine insulin may be a model process for the production of other human insulin analogs.