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Characterization and Enhanced Production of Enterocin HJ35 by Enterococcus faecium HJ35 Isolated from Human Skin

  • Yoon Yoh Chang;Park Hye Jung;Lee Na-Kyoung;Paik Hyun-Dong
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.4
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    • pp.296-303
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    • 2005
  • A strain named as HJ35 was isolated from the skin of sixty-five men and fourteen women for acne therapy, in order to find an effective antimicrobial agent against Propionibacterium acnes. Isolate HJ35 was identified as Enterococcus faecium based on 16 rDNA sequence and produced enterocin HJ35 having antimicrobial activities against most lactic acid bacteria, En­terococcus spp., Staphylococcus aureus, S. epidermidis, Clostridium perfringens, some bacilli, Mi­crococcus flavus, Listeria monocytogenes, L. ivanovii, Escherichia coli, Pseudomonas fluorescens and Propionibacterium acnes, in the modified well diffusion method. Especially, enterocin HJ35 showed a bactericidal activity against Propionibacterium acnes P1. The antimicrobial activity of enterocin HJ35 was disappeared completely with the use of protease XIV. But enterocin HJ35 activity is very stable at high temperature (up to $100^{\circ}C$ for 30 min), in wide range of pH (3.0${\~}$9.0), and by treatment with organic solvents. The apparent molecular mass of enterocin HJ35 was estimated to be approximately 4${\~}$4.5 kDa on detection of its bactericidal activity after SDS-PAGE. In batch fermentation of E. faecium HJ35, enterocin HJ35 was produced at the mid­log growth phase, and its maximum production was obtained up to 2,300 AU/mL at the late stationary phase. By employing fed-batch fermentation, the enhanced production of enterocin HJ35 was achieved up to 12,800 AU/mL by feeding with 10 g/L glucose or 6 g/L lactate.

Dewatering and Settling Characteristics of Digested Sludge from the Anaerobic Sequencing Batch Reactor for Treatment of Nightsoil (혐기성 연속회분식 공정을 이용한 분뇨처리시 소화슬러지의 침강 및 탈수특성)

  • Lee, Jun-Geol;Jang, Duk;Heo, Jun-Mu;Son, Boo-Soon;Jang, Bong-Ki;Park, Jong-An
    • Journal of Environmental Science International
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    • v.7 no.6
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    • pp.867-873
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    • 1998
  • Laboratory study was conducted to provide basic data for operating anaerobic sequencing batch reactor(ASBR) process for treatment of nightsoil. The experiments were concerned with digestion characteristics, settleabiltity and dewaterability of digested sludge in ASBR system. Completely-mixed dally-fed control reactor without solid-liquid separation step was also operated to evaluate the baseline performance since the nature of nightsoil was changed with time. In all case, gas production from the ASBR shows 1.3 to 1.44 times higher than that from control, in spite of almost similar trend in organics removal. During thickening period, remarkable decrease in surface settling velocity was observed at the ASBRs compared with the control. In case of the ASBR run, flotation of mixed digested sludge was not occurred. Also, ultimate thickened volume of ASBRs increased 1.2~1.5 times compared with control. Dewaterability of digested sludge without conditioning decreased when the completely-mixed daily-fed reactor for ASBR run was converted to the ASBR. However, improvement of dewaterability of digested sludge from the ASBRs was observed as a result of addition of FeCl$_3$ to digested sludge for conditioning.

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Optimization of D-\beta-hydroxybutyric Acid Fermentation Using a Mutant of Candida Rugosa IFO0750 (Candida rugosa 변아주를 이용한 D-\beta-Hydroxybutyric Acid 발효공정의 최적화)

  • 경수현;신철수
    • Microbiology and Biotechnology Letters
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    • v.28 no.6
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    • pp.355-360
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    • 2000
  • A UVmutant of Candida rugosa IF00750 was made and used to convert butYlic acid to D-$\beta$-hydroxybutyric acid(D-$\beta$-HBA). Major regulating factors for D-$\beta$-HBA fennentation were investigated via chemostat analyses. The maximum specific productivity was achieved at a specific growth rate of $0.06h^{-1}$ where the glucose and butyric acid concentrations in the fermentor were 10 g/L and 8.7 g/L. respectively. A fed-batch fennentation was performed with maintenance of the optimum glucose and butyric acid concentrations. The D-$\beta$-HBA concentration after 120 h of cultivation reached 12.4 g/L, which was 4.7 times greater illan the concentration obtained by batch fermentation.

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High-Level Expression of Aspergillus ficuum Acetyl Xylan Esterase Gene in Pichia pastoris, (Pichia pastoris에서 Aspergillus ficuum 유래 Acetyl Xylan Esterase 유전자의 과발현)

  • 임재명;김성구;박승문;남수완
    • Microbiology and Biotechnology Letters
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    • v.30 no.4
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    • pp.305-311
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    • 2002
  • Acetyl xylan esterase gene (AXE) from Aspergillus ficuum was cloned and its Pichia expression plasmid, pPICZ$\alpha$C-AXE (4.6 kb), was constructed, in which the AXE gene was under the control of the AOXI promoter and connected downstream of mating factor u-1 signal sequence. The plasmid linearized by Sacl was integrated into the 5'AOXI region of the chromosomal DNA of P. pastoris. In the flask batch culture of P. pastoris transformant on methanol medium, the cell concentration and total AXEase activity reached at 6.0 g-dry cell weight/1 and 77 unit/ml after 36 h cultivation, respectively. In the fed-batch culture employing the optimized methanol and histidine feeding strategy, the cell concentration and total AXEase activity were significantly increased to about 97 g-dry cell weight/1 and 930 unit/ml. Most of AXEase activity (>90%) was found in the extracellular medium and the majority of extracellular protein (>80%) was AXEase enzyme (33.5 kDa). This result means that about 9.8 g/1 of AXEase protein was produced in the extracellular medium.

Enhancement of BDNF Production by Co-cultivation of Human Neuroblastoma and Fibroblast Cells

  • Hong, Jong-Soo;Oh, Se-Jong;Kim, Sun-Hee;Park, Kwon-Tae;Cho, Jin-Sang;Park, Kyung-You;Lee, Jin-Ha;Lee, Hyeon-Yong
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.3 no.2
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    • pp.51-54
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    • 1998
  • It has been proved that co-cultivation of human neroblastoma cells and human fibroblast cells can enhance nerve cell growth and the production of BDNF in perfusion cultivation. In batch co-cultivation, maximum cell density was increased up to 1.76${\times}$106 viable cells/mL from 9${\times}$105 viable cells/mL of only neuroblastoma cell culture. The growth of neuroblastoma cells was greatly improved by culturing both nerve and fibroblast cells in a perfusion process, maintaining 1.5${\times}$106 viable cells/mL, which was much higher than that form fed-batch cultivation. The nerve cell growth was greatly enhance in both fed-batch and perfusion cultivations while the growth of fibroblast cells was not. It strongly implies that the factors secreted from human fibrobast cells and/or the environments of co-culture system can enhance both cell growth and BDNF secretion. Specific BDNF production rate was not enhanced in co-cultures; however, the production period was increased as the cell growth was lengthened in the co-culture case. Competitive growth between nerve cells and fibroblast cells was not observed in all cases, showing no changes of fibroblast cell growth and only enhancement of the neuroblastoma cell growth and overall BDNF production. It was also found that the perfusion cultivation was the most appropriate process for cultivating two cell lines simultaneously in a bioreactor.

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Cloning, Characterization of Pichia etchellsii $\beta-Glucosidase$ II and Effect of Media Composition and Feeding Strategy on its Production in a Bioreactor

  • Sethi Benu;Jain Monika;Chowdhary Manish;Soni Yogesh;Bhatia Yukti;Sahai Vikram;Mishra Saroj
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.1
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    • pp.43-51
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    • 2002
  • The cloning and expression of $\beta-glucosidase$ II, encoded by the gene ${\beta}glu2$, from thermotolerant yeast Pichia etchellsii into Escherichia coli is described. Cloning of the 7.3 kb BamHI/SalI yeast insert containing ${\beta}glu2$ in pUC18, which allowed for reverse orientation of the insert, resulted in better enzyme expression. Transformation of this plasmid into E. coli JM109 resulted in accumulation of the enzyme in periplasmic space. At $50^{\circ}C$, the highest hydrolytic activity of 1686 IU/g protein was obtained on sophorose. Batch and fed-batch techniques were employed for enzyme production in a 14 L bioreactor. Exponential feeding rates were determined from mass balance equations and these were employed to control specific growth rate and in turn maximize cell growth and enzyme production. Media optimization coupled with this strategy resulted in increased enzyme units of 1.2 kU/L at a stabilized growth rate of $0.14\;h^{-l}$. Increased enzyme production in bioreactor was accompanied by formation of inclusion bodies.

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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Optimization of Fermentation Process for Acetic Acid Production (초산 생성을 위한 발효공정의 최적화)

  • Shin, Jin-A;Oh, Nam-Soon
    • Food Engineering Progress
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    • v.14 no.3
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    • pp.217-221
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    • 2010
  • Various conditions of acetic acid fermentation by Acetobacter aceti B20 strain were investigated and evaluated to optimize the fermentative production of acetic acid. The effects of the initial ethanol concentration on growth and acid productivity in a flask and fermentor were also studied. The growth of A. aceti B20 strain was inhibited as the concentration of ethanol increased. However, the highest total acidity and fermentation yield were 5.34% and 56.1%, respectively when the initial concentration of ethanol was 7% in the batch fermentation. Although the concentration of initial glucose influenced the growth rate of B20 strain, it did not influence the total acidity in the flask culture. When the agitation speed increased, the growth, total acidity and fermentation yield were all improved. In fed-batch fermentation, total acidities and fermentation yields were 7.14-8.76% and 39.1-53.0%, respectively, and their values mostly depended on the feeding methods.

Overproduction of Xanthophyll Pigment in Flavobacterium sp. JSWR-1 under Optimized Culture Conditions

  • Jegadeesh Raman;Young-Joon Ko;Jeong-Seon Kim;Da-Hye Kim;Soo-Jin Kim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.3
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    • pp.710-724
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    • 2024
  • Flavobacterium can synthesize xanthophyll, particularly the pigment zeaxanthin, which has significant economic value in nutrition and pharmaceuticals. Recently, the use of carotenoid biosynthesis by bacteria and yeast fermentation technology has shown to be very efficient and offers significant advantages in large-scale production, cost-effectiveness, and safety. In the present study, JSWR-1 strain capable of producing xanthophyll pigment was isolated from a freshwater reservoir in Wanju-gun, Republic of Korea. Based on the morphological, physiological, and molecular characteristics, JSWR-1 classified as belonging to the Flavobacterium species. The bacterium is strictly aerobic, Gram-negative, rod-shaped, and psychrophilic. The completed genome sequence of the strain Flavobacterium sp. JSWR-1 is predicted to be a single circular 3,425,829-bp chromosome with a G+C content of 35.2% and 2,941 protein-coding genes. The optimization of carotenoid production was achieved by small-scale cultivation, resulting in zeaxanthin being identified as the predominant carotenoid pigment. The enhancement of zeaxanthin biosynthesis by applying different light-irradiation, variations in pH and temperature, and adding carbon and nitrogen supplies to the growth medium. A significant increase in intracellular zeaxanthin concentrations was also recorded during fed-batch fermentation achieving a maximum of 16.69 ± 0.71 mg/l, corresponding to a product yield of 4.05 ± 0.15 mg zeaxanthin per gram cell dry weight. Batch and fed-batch culture extracts exhibit significant antioxidant activity. The results demonstrated that the JSWR-1 strain can potentially serve as a source for zeaxanthin biosynthesis.

The Role of Primary Clarifier in Biological Processes for Nutrient Removal (생물학적 질소·인제거 공정에서 일차 침전지의 영향)

  • Whang, Gye-Dae;Kim, Tae-Kyung
    • Journal of Korean Society on Water Environment
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    • v.23 no.1
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    • pp.19-26
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    • 2007
  • The lab-scale BNR processes fed with Municipal Wastewater Before or After Primary Clarifier (MWBPC or MWAPC) were operated to observe the behavior of particle organic matter in terms of nitrification and denitrification efficiency. As a result of the fractionation of the COD from MWBPC or MWAPC using an aerobic respirometric serum bottle reactor, the total mass of biodegradable organic matter from MWBPC is about 52% greater than the mass from MWAPC. Batch reactors were operated to observe the effect of the Particulate Organic Matter (POM) on substrate utilization for denitrification. Although the consumption of POM for denitrification was observed, the increment of the Specific Denitrification Rate (SDNR) was not great. In terms of the effect of POM on nitrification at different HRTs, activate sludge reactors were operated to determine the optimal HRT when MWBPC and MWAPC were fed relatively. All reactors showed a great organic matter removal efficiency. Reactors fed with MWAPC had obtained the nitrification efficiency above 90% when the HRT of 4 hr, at least, was maintained, while reactors fed with MWBPC had same efficiency when the HRT longer than 5 hr was kept. Three parallel $A^2/O$ systems fed with MWBPC or MWAPC relatively were operated to investigate the effects of POM on BNR processes with varying the HRT of an anoxic reactor. For all systems, the efficiency of organic matter removal and denitrification, respectively, was great and about the same. In case of denitrification efficiency, system with MWAPC had 1.5% lower than system with MWBPC at the same HRT of anoxic reactor of 2 hr, and the increasing the HRT of the anoxic reactor by 1 hr in systems fed with MWBPC resulted in a 3.5% increment. The denitrification rate was similar while the consumption of organic matter in systems fed with MWBPC was higher than system fed with MWBPC. It suggests that POM in MWBPC was not be used significantly as a substrate for denitrification in system with the HRT of 3 hr of an anoxic reactor.