• Title/Summary/Keyword: fermentor

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Isolation and Identification of Linear Alkylbenzene Sulfonate Degrading Bacteria (Linear Alkylbenzene Sulfonate 분해세균의 분리 및 동정)

  • Lee, Ki-Moo;Choi, Woo-Young
    • Korean Journal of Agricultural Science
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    • v.21 no.1
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    • pp.60-66
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    • 1994
  • Among the various bacterial isolates from municipal sewages which utilized linear alkylbenzene sulfonate (LAS) as a sole source of carbon. 3 potent strains - KL-3, SH-2 and EN-1 - were selected. The strains were classified: KL-3 as a strain belong to the genus Klebsiella; SH-2 Shigella; and EN-1 Enterobacter, respectively. They were grown in a broth containing 200 ppm of LAS, using a laboratory scale fermentor: the bacterial growth reached stationary phase after 2 days with a maximum viability of $10^8cfu/mL$ of the culture; initial rates of LAS degradation were high during the first 24 hours of cultivation (KL-3 and SH-2, approx. 50%; EN-1, 20%); after 1 day a lag period of about 24 hours was observed for all the strains, and thereafter break-down proceeded rapidly; final rates after 7 days were approximately 85% by KL-3, 82% by SH-2 and 75% by EN-1. Adsorption of LAS by the bacterial cell mass was high for the strain SH-2, as Freundlich equation: Y= 0.030X + 0.95 was calculated.

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Fed-batch Cultivation of Escherichia coli YK537 (pAET-8) for Production of phoA Promoter-controlled Human Epidermal Growth Factor

  • Wang Yonggang;Du Peng;Gan Renbao;Li Zhimin;Ye Qin
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.2
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    • pp.149-154
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    • 2005
  • Secretion of the expressed heterologous proteins can reduce the stress to the host cells and is beneficial to their recovery and purification. In this study, fed-batch cultures of Escherichia coli YK537 (pAET-8) were conducted in a 5-L fermentor for the secretory production of human epidermal growth factor (hEGF) whose expression was under the control of alkaline phosphatase promoter. The effects of feeding of glucose and complex nitrogen sources on hEGF production were investigated. When the fed-batch culture was conducted in a chemically de-fined medium, the cell density was 9.68 g/L and the secreted hEGF was 44.7 mg/L in a period of 60 h. When a complex medium was used and glucose was added in pH-stat mode, the secreted hEGF was improved to 345 mg/L. When the culture was fed with glucose at a constant specific rate of $0.25\;gg^{-1}h^{-1}$, hEGF reached 514 mg/L. The effects of adding a solution containing yeast extract and tryptone were further studied. Different rate of the nitrogen source feeding resulted in different levels of phosphate and acetic acid formation, thus affected hEGF expression. At the optimal feeding rate, hEGF production achieved 686 mg/L.

Purification and Characterization of the Antitumor Antibiotic from Streptomyces sp. YBE-316 (Streptomyces sp. YBE-316이 생산하는 항암성 항생물질의 정제 및 특성)

  • Park, Jae-Hong;Hahm, Byoung-Kwon;Bai, Dong-Hoon;Yu, Ju-Hyun
    • Microbiology and Biotechnology Letters
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    • v.23 no.3
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    • pp.329-336
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    • 1995
  • For the development of new antitumor antibiotics produced by microorganisms, Streptomyces sp. YBE-316 was isolated from soil. The productivity of the antitumor antibiotic from Streptomyces sp. YBE-316 gradually increased after 60 hours, and was maximum after 100 hours after inoculation in growth medium (2.0% sucrose, 1.0% soybean meal, 0.1% K$_{2}$HPO$_{4}$, pH 7.0) at 30$\circ$C, 150 rpm, 5 NL/min by 30 l jar fermentor. This antitumor antibiotic was present only in mycelium, and stable in pH 5.0-10.0 for 20 minutes at 100$\circ$C. Antitumor and antibiotic activities were maintained at neutral pH, and heat stability was low. This antitumor antibiotic was soluble in methanol and ethanol, and insoluble in water, ethyl acetate, chloroform, and n-hexane. This antitumor antibiotic was sequentially purified by acetone extraction from mycelium, butanol extraction, and silica gel column chromatography. Antitumor activity was low against most tested cell lines, but antibiotic activity was high and low against yeasts and bacteria, respectivelv. The visualization test showed that this antitumor antibiotic had higher hydroxyl, ketone, amino, carboxyl groups, and sugar(s) in its structure. Instrumental analyses showed that this antitumor antibiotic was a pentaene in polyene class antibiotics. In pentaene class antibiotics, this was considered as an eurocidin or capacidin type antibiotics. The molecular weight of this antitumor antibiotic was higher than 683.0 daltons, and this antitumor antibiotic might be glycosylated by other sugar(s), instead of mycosamine or perosamine, an amino sugar.

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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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Production Conditions of Two Polysaccharides from Marine Bacterium Zoogloea sp. (해양세균 Zoogloea sp.로부터 두 종류의 polysaccharide 생산조건)

  • 장명웅;강양순
    • KSBB Journal
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    • v.10 no.5
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    • pp.518-524
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    • 1995
  • Marine bacterium, as a microbial source producing polysaccharides, was newly isolated from the eastern and western sea of Korea and was identified as Zoogloea sp. (KCCM 10036). It produced two different types of polysaccharides, especially: WSP (water-soluble polysaccharide) and CBP (cell-bound polysaccharide). The former was isolated from the supernatant of centrifuged broth by acetone precipitation, and the latter was isolated from the pellet by acetone and CPC (cetylpyrldinium chloride) precipitation. The productivity of polysaccharides were increased with the addition of promoting agents such as biotin, ampicillin and surfactant. After batch fermenting, the productivity of WSP and CBP were reached to maximum values of $9.Og/\ell$, $2.5g/\ell$ in the culture medium containing 1% of glucose as a carbon source.

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Enhanced Production of C30 Carotenoid 4,4'-Diaponeurosporene by Optimizing Culture Conditions of Lactiplantibacillus plantarum subsp. plantarum KCCP11226T

  • Siziya, Inonge Noni;Yoon, Deok Jun;Kim, Mibang;Seo, Myung-Ji
    • Journal of Microbiology and Biotechnology
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    • v.32 no.7
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    • pp.892-901
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    • 2022
  • The rising demand for carotenoids can be met by microbial biosynthesis as a promising alternative to chemical synthesis and plant extraction. Several species of lactic acid bacteria (LAB) specifically produce C30 carotenoids and offer the added probiotic benefit of improved gut health and protection against chronic conditions. In this study, the recently characterized Lactiplantibacillus plantarum subsp. plantarum KCCP11226T produced the rare C30 carotenoid, 4,4'-diaponeurosporene, and its yield was optimized for industrial production. The one-factor-at-a-time (OFAT) method was used to screen carbon and nitrogen sources, while the abiotic stresses of temperature, pH, and salinity, were evaluated for their effects on 4,4'-diaponeurosporene production. Lactose and beef extract were ideal for optimal carotenoid production at 25℃ incubation in pH 7.0 medium with no salt. The main factors influencing 4,4'-diaponeurosporene yields, namely lactose level, beef extract concentration and initial pH, were enhanced using the Box-Behnken design under response surface methodology (RSM). Compared to commercial MRS medium, there was a 3.3-fold increase in carotenoid production in the optimized conditions of 15% lactose, 8.3% beef extract and initial pH of 6.9, producing a 4,4'-diaponeurosporene concentration of 0.033 A470/ml. To substantiate upscaling for industrial application, the optimal aeration rate in a 5 L fermentor was 0.3 vvm. This resulted in a further 3.8-fold increase in 4,4'-diaponeurosporene production, with a concentration of 0.042 A470/ml, compared to the flask-scale cultivation in commercial MRS medium. The present work confirms the optimization and scale-up feasibility of enhanced 4,4'-diaponeurosporene production by L. plantarum subsp. plantarum KCCP11226T.

Lipase Production by Limtongozyma siamensis, a Novel Lipase Producer and Lipid Accumulating Yeast

  • Varunya Sakpuntoon;Savitree Limtong;Nantana Srisuk
    • Journal of Microbiology and Biotechnology
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    • v.33 no.11
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    • pp.1531-1541
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    • 2023
  • Lipase is a well-known and highly in-demand enzyme. During the last decade, several lipase optimization studies have been reported. However, production costs have always been a bottleneck for commercial-scale microbial enzyme production. This research aimed to optimize the conditions for lipase production by Limtongozyma siamensis DMKU-WBL1-3 via a One-Factor-At-a-Time (OFAT) approach combined with statistical methods while using a low-cost substrate. Results suggest that low-cost substrates can be substituted for all media components. An optimal medium was found, using response surface methodology (RSM) and central composite design (CCD), to consist of 0.50% (w/v) sweet whey, 0.40% (w/v) yeast extract (food grade), and 2.50% (v/v) palm oil with the medium pH adjusted to 4 under shaking flask cultivation. From an economic point of view, this work was successful in reducing production costs while increasing lipase productivity. The medium costs were reduced by 87.5% of the original cost while lipase activity was increased by nearly 6-fold. Moreover, lipase production was further studied in a 2-L stirred-tank fermentor. Its activity was 1,055.6 ± 0.0 U/ml when aeration and agitation rates were adjusted to 1 vvm and 170 rpm, respectively. Interestingly, under this optimal lipase production, the yeast showed accumulated lipids inside the cells. The primary fatty acid is a monounsaturated fatty acid (MUFA) that is typically linked to health benefits. This study hence reveals promising lipase production and lipid accumulation by L. siamensis DMKU-WBL1-3 that are worthy of further study.

Production and Characterization of Extracellular Polysaccharide Produced by Pseudomonas sp. GP32 (Pseudomonas sp. GP32에 의해 생산된 세포 외 다당류의 생산 및 특성)

  • Lee, Myoung Eun;Lee, Hyun Don;Suh, Hyun-Hyo
    • Journal of Life Science
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    • v.25 no.9
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    • pp.1027-1035
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    • 2015
  • A strain GP32 which produces a highly viscous extracellular polysaccharide was conducted with soil samples and identified as Pseudomonas species. The culture flask conditions for the production of extracellular polysaccharide by Pseudomonas sp. GP32 were investigated. The most suitable carbon and nitrogen source for extracellular polysaccharide production were galactose and (NH4)2SO4. The optimum carbon/nitrogen ratio for the production of extracellular polysaccharide was around 50. The optimum pH and temperature for extracellular polysaccharide production was 7.5 and 32℃, respectively. In batch fermentation using a jar fermentor, the highest extracellular polysaccharide content (15.7 g/l) was obtained after 70 hr of cultivation. The extracellular polysaccharide produced by Pseudomonas sp. GP32 (designated Biopol32) was purified by ethanol precipitation, cetylpyridinium chloride (CPC) precipitation, and gel permeation chromatography. Biopol32, which has an estimated molecular weight of over 3×107 datons, is a novel polysaccharide derived from sugar components consisting of galactose, glucose, gulcouronic acid and galactouronic acid in an approximate molar ratio of 1.85 : 3.24 : 1.00 : 1.42. The solution of Biopol32 showed non-Newtonian characteristics. The viscosity of Biopol32 exhibited appeared to be higher at all concentration compared to that of zooglan from Zoogloea ramigera. An analysis of the flocculating efficiency of Biopol32 in industry wastewater (food, textile, and paper wastewater) revealed chemical oxygen demand (COD) reduction rates 58.4-67.3% and suspended solid (SS) removal rates 82.6-91.3%. Based on these results, Biopol32 is a possible candidate for industrial applications such as wastewater treatment.

Isolation and Mycelial Cultivation Submerged of Phellinus sp. (Phellinus sp.의 분리 및 균사체의 액체배양)

  • Kang, Tae-Su;Lee, Dong-Gi;Lee, Shin-Young
    • The Korean Journal of Mycology
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    • v.25 no.4 s.83
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    • pp.257-267
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    • 1997
  • Fruit bodies similar to the Phellinus sp. residing on the mulberry were collected at Yang-yang in Kang-won-do province and one strain of Phellinus sp. was isolated from the fruit bodies. For mass production of the isolated mycelia in a submerged culture, the culture conditions, medium composition, and the effect of various culture systems on the mycelial growth, were investigated. The morphological characteristics of the fruit body were as follows: covered with blackish to black and rough, lower surface with yellowish-brown to dull-brown and smooth, 5-7 cm thick and hard woody. Also, the pure cultured mycelia showed yellowish-brown color, capability of purplish-brown pigment production on the PDA plate media, no-formation of clamp-connection, much binding branch, and enzyme activities such as laccase, tyrosinase and peroxidase. Therefore, pure cultured strain was identified to be Phellinus sp. In the flask culture, the optimum culture conditions for the mycelial production were obtained after cultivation of 8 days at inoculum level of 5%(v/v), media volume of 70 mL, 150 rpm, initial pH 6, and temperature of $30^{\circ}C$. Optimum medium composition from the response surface analysis were determined to be glucose 12.12 g/L, sucrose 12.12 g/L, yeast extract 11.15 g/L, malt extract 11.15 g/L, $KH_2PO_4$ 0.855 g/L and $CaCl_2$ 0.855 g/L. The production of the mycelia after 4 and 8 days of cultivation was 1.95 and 9.89 g/L, respectively. The maximum specific growth rate and productivity were $0.020\;hr^{-1}$ and 1.25 g/L/day, respectively. Among the three different culture systems for the growth of mycelia, the maximum mycelial dry weight of 7.5 g/L was obtained after cultivation of 4 days in the air-lift fermentor under aeration rate of 2.5 vvm. The maximum specific growth rate and productivity were $0.033\;hr^{-1}$ and 1.9 g/L/day, respectively, which were about 1.7 and 4.2 times higher than those of flask culture.

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Optimization of the Acetic Acid Fermentation Condition of Apple Juice (사과식초 제조를 위한 사과주스의 초산발효 최적화)

  • Kang, Bok-Hee;Shin, Eun-Jeong;Lee, Sang-Han;Lee, Dong-Sun;Hur, Sang-Sun;Shin, Kee-Sun;Kim, Seong-Ho;Son, Seok-Min;Lee, Jin-Man
    • Food Science and Preservation
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    • v.18 no.6
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    • pp.980-985
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    • 2011
  • This study was conducted to determine the acetic-acid fermentation properties of apple juice (initial alcohol content, apple juice concentration, acetic-acid content, and inoculum size) in flask scale. At the acetic-acid fermentation of apple juice with 3, 5, 7, and 9% initial alcohol content, the maximum acidity after 10-day fermentation was 5.88% when the initial alcohol content was 5%. The acetic-acid fermentation did not proceed normally when the initial alcohol content was 9%. When the initial Brix was $1^{\circ}$, the acidity gradually increased, and the acidity after 12-day acetic-acid fermentation was 4.48%. Above 4% acidity was attained faster when the apple juice concentration was 5 and 10 $^{\circ}Brix$ than when it was 1 and 14 $^{\circ}Brix$. When the initial acidity was 1% or above (0.3, 0.5, 1.0, and 2.0%), the acetic-acid fermentation proceeded normally. The acetic-acid fermentation also proceeded normally when the inoculum sizes were 10 and 15%, and the acidity after eight-day acetic-acid fermentation was 5.60 and 6.05%, respectively. Therefore, the following were considered the optimal acetic-acid fermentation conditions for apple cider vinegar: 5% initial alcohol content, 5 $^{\circ}Brix$ or above apple juice concentration, 1.0% or above initial acidity, and 10% or above inoculum size. Apple cider vinegar with above 5% acidity can be produced within 48 h under the following acetic-acid fermentation conditions: 7% initial alcohol content, about 1% initial acidity, and 10% inoculum volume at $30^{\circ}C$, 30 rpm, and 1.0 vvm, using 14 $^{\circ}Brix$ apple juice in a mini-jar fermentor as a pre-step for industrial-scale adaptation.