• Title/Summary/Keyword: Entrapping

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Studies on Microbial Penicillin Amidase (Part 6) Immobilization of Penicillin Amidase from Bacillus megaterium by Adsorption and Acrylamide Gel Entrappment (미생물 페니실린 아미다제에 관한 연구 (제 6 보) 흡착효소의 아크릴아마이드젤 포괄방법에 의한 Bacillus megaterium의 변이주가 생산하는 페니실린 아미다제의 고정화에 관한 연구)

  • Seong, Baik-Lin;Son, Hyeung-Jin;Mheen, Tae-Ick;Moon H. Han
    • Microbiology and Biotechnology Letters
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    • v.9 no.4
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    • pp.197-205
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    • 1981
  • Penicillin amidase of Bacillus megaterium was recovered from the fermentation broth by adsorption on celite and immobilized by entrapping the adsorbed enzyme in acrylamide gel. The operational stability in column reactor was greatly increased by entrappment as compared with that of without entrappment. The optimum pH of the immobilized enzyme was 8.7 with broader activity profile than that of the free enzyme, while the most stable pH range appeared to be between pH 7.5 and 8.0. The optimum temperature was shifted to 5$0^{\circ}C$ from 45$^{\circ}C$ for the soluble enzyme. The values of Km and the inhibition constants for 6-APA( $K_{ia}$ ) and phenylacetic acid ( $K_{ip}$ ), were 4.55 mM, 36.5mM, and 10.5mM, respectively. No significant internal pore diffusion limitation was found since the value of effectiveness factor was 0.95. The operational half life in a column reactor at pH 8.0 was 6.8 days at 4$0^{\circ}C$ and 47 days at 3$0^{\circ}C$, whereas that of without entrappment was only 1 day and 4 days, respectively. The performance of a batch and a column reactor was also discussed with respect to the productivity. The results demonstrated that the entrappment of an adsorbed enzyme for the enhancement of the operational stability of the immobilized enzyme was useful especially when an extracellular enzyme was used.

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Use of Bacteriocin Produced by Lactococcus sp. CU216 with pH Sensitive Liposome Entrapment (Lactococcus sp. CU216이 생산하는 박테리오신을 함유한 pH Sensitive Liposome의 응용)

  • 박성수;김명희;한경식;오세종
    • Food Science of Animal Resources
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    • v.24 no.1
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    • pp.97-102
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    • 2004
  • The objective of this study was to control Kimchi fermentation using pH sensitive bacteriocin entrapping liposome(bacteriocin-liposome). The liposomes were prepared by the reverse-phase evaporation method from a mixture of DPPC(dipalmitoyl phosphatidylcholine, DPPE(dipalmitoyl phosphatidylethanolamine), DOPC(dioleoyl phosphatidylcholine) and cholesterol in a molar ration of 4:2:1:4. The bacteriocin-liposome was disruptured at pH 4 of buffer and was stable at alkaline pHs(6 and 7). Irrespective of the addition of the bacteriocin-liposomes, the pH of every Kimchi sample decreased to 5 during 5 days storage at 5$^{\circ}C$. Kimchi samples treated with bacteriocin-liposomes maintained pH 4 or higher, while Kimchi samples not treated with bacteriocin-liposomes exhibited pH 3.58 or lower. In general, the pH of Kimchi samples stored at 20$^{\circ}C$ decreased faster, compared to that of Kimchi samples stored at 5$^{\circ}C$. The pH of Kimchi samples treated with the bacteriocin-liposomes was 3.9 during 90 days storage, while that of the samples not treated with the bacteriocin-liposomes was 3.68 and 3.32 during 30 days and 90 days storages, respectively. Lactic acid bacteria in Kimchi treated with the bacteriocin-liposome grew relatively slow at 5$^{\circ}C$. The viable cell number of lactic acid bacteria increased up to 4${\times}$10$\^$7/ cells/ml and then decreased to 8${\times}$10$\^$6/ cells/ml during 90 days storage at 5$^{\circ}C$.

Studies on Mass Production of Intracellularly-Produced Secondary Metabolite, Cyclosporin A by Use of Immobilized Fungal Cells in Stirred-Tank Immobilized Perfusion Reactor System(IPRS) (교반식 perfusion 생물반응기(IPRS)에서 고밀도 고정상 곰팡이 세포를 이용한 세포내 축적 이차대사산물인 Cyclosporin A 대량생산에 관한 연구)

  • 전계택;이태호장용근
    • KSBB Journal
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    • v.11 no.1
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    • pp.22-29
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    • 1996
  • Immobilized bioprocess was carried out for continuous production of cyclosporin A (CyA) produced intracellularly as a secondary metabolite by a filamentous fungus, Tolypocladium inflatum. Immobilization procedure for entrapping conidiospores of the producer was significantly simplified by use of a modified immobilization technique. A newly-designed immobilized perfusion reactor system (IPRS) showed good process benefits as demonstrated by the role of the high density immobilized cells as an efficient biomass generator, continuously supplying highly active CyA-producing free cells (1.0g/$\ell$/hr) even at very high dilution rate ($0.1hr^{-1}$). IPRS bioprocess was possible since efficient decantor system developed in our laboratory separated the sloughed-off free cells from the immobilized biomass effectively, thus overcoming wash-out phenomenon frequently encountered in continuous free cell cultures. Furthermore the released-free cells remaining in the bulk solution did not appear to cause substrate mass transfer limitation which was often experienced in suspended mycelial fungal cell fermentations. The primary reason for this was that the suspension broth of the IPRS mainly consisted of roundshaped short mycelial fragments and conidiospores, still remaining Newtonian even at high cell density. In parallel with IPRS bioprocess development, other key factors to be considered necessarily for significant increase in CyA productivity would be strain improvement and medium optimization for the immobilized cells.

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Production of Nucleotide by Immobilized Cell (고정화 미생물에 의한 뉴크레오타이드 생산)

  • CHO Jung-Il;JUNG Sung-Won
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.24 no.2
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    • pp.111-116
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    • 1991
  • The effective p.eduction of 5'-GMP(5'-Guanylic acid) by enzymatic conversion of 5'-XMP(5'-Xanthyic acid) was investigated. The Iyophilized Brevibacterium ammoniagenes ATCC 19216 which were used as the XHP aminase source, was immobilized by entrapping in K-carrageenan, agar, polyacrylamide or Ca-alginate. $3\%$ K-carrageenan was selected as the most suitable matrix. In the production of 5'-GMP using the free cells of 3. ammoniagenes ATCC 19216, the optimum conditions were $42^{\circ}C$, PH 7.0, 100mg/ml glucose, 120mg/ml cell ,8mg/ml $MgSO_4\cdot7H_2O$, 5mg/ml POESA, 5mg/ml phytic acid. Under the conditions, $94.5\%$ of 5'-GMP was converted within 8 hours. In the production of 5'-GMP using the immobilized whole cells of B. ammoniagenes ATCC 19216, the optimum conditions were $37^{\circ}C$, pH 7.5, 50mg/ml glucose, 1mg/ml $KH_2PO_4$, 10mg/ml phytic acid, 60mg/ml cell, 8mg/ml $MgSO_4\;\cdot\;7H_2O$, 5mg/ml POESA. Under the conditions, $64.7\%$ of 5'-GMP was converted within 40 hours.

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Performance Evaluation of Biofuel cell using Benzoquinone Entrapped Polyethyleneimine-Carbon nanotube supporter Based Enzymatic Catalyst (벤조퀴논 포집 폴리에틸렌이민-탄소나노튜브 지지체 기반 효소촉매의 바이오연료전지로서의 성능평가)

  • Ahn, Yeonjoo;Chung, Yongjin;Kwon, Yongchai
    • Korean Chemical Engineering Research
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    • v.55 no.2
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    • pp.258-263
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    • 2017
  • In this study, we synthesized biocatalyst consisting of glucose oxidase (GOx), polyethyleneimine (PEI) and carbon nanotube (CNT) with addition of p-benzoquinone (BQ) that was considered anodic catalysts of enzymatic biofuel cell (EBC). For doing this, PEI/CNT supporter was bonded with BQ by physical entrapping method stemmed from electrostatic attractive force ([BQ/PEI]/CNT). In turn, GOx moiety was further immobilized on the [BQ/PEI]/CNT to form GOx/[BQ/PEI]/CNT catalyst. This catalyst has a special advantage in that the BQ that has been usually dissolved into electrolyte was immobilized on supporter. According to the electrochemical analysis, maximum current density of the GOx/[BQ/PEI]/CNT catalyst was 1.9 fold better than that of the catalyst that did not entrap BQ with the value of $34.16{\mu}A/cm^2$, verifying that catalytic activity of the catalyst was enhanced by adoption of BQ. Also, when it was used as anodic catalyst of the EBC, its maximum power density was 1.2 fold better than that of EBC using the catalyst that did not entrap BQ with the value of $0.91mW/cm^2$. Based on such results, it turned out that the GOx/[BQ/PEI]/CNT catalyst was promising and viable as anodic catalyst of EBC.

Immobilization of Zygosaccharomyces rouxii (Zygosaccharomyces rouxii의 고정화(固定化))

  • Park, Se Jeong;Park, Yoon Joong
    • Korean Journal of Agricultural Science
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    • v.14 no.1
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    • pp.156-163
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    • 1987
  • In these experiment, the conditions of entrapping immobilization of Zygosaccharomyces rouxii that participate in the soysauce brewing are investigated. And carried out the fermentation and aging test by immobilized Zygosaccharomyces rouxii with the hydrolyzed solution prepared from soysauce koji. The results obtained were as follows: 1. Immobilizing conditions of Zygosaccharomyces rouxii. 1) When the concentration of Na-alginate solution is 2.0-2.5%, the bead formation was very good. And the concentration of Na-alginate solution not influenced on the fermentation activity of immobilized Zygosaccharomyces rouxii. 2) Effect of ratio of the precultured Zygosaccharomyces rouxii solution and Na-alginate solution on the fermentation activity of immobilized Zygosaccharomyces rouxii was not highly recognized. But if the ratio of precultured Zygosaccharomyces rouxii solution increased, the fermentation activity of immobilized Zygosaccharomyces rouxii was slightly high. 3) The fermentation activity of immobilized Zygosaccharomyces rouxii that grew over 36hrs was higher than that grew below 24hrs. 4) Increasing the ratio of immobilized Zygosaccharomyces rouxii gel to the fermentative medium, the fermentation activity of Zygosaccharomyces rouxii was higher. 2. The fermentation test by immobilized Zygosaccharomyces rouxii with the hydrolyzed solution of soysauce koji. 1) When fermented for about 96 hrs, the alcoholic fermentation almost stopped and alcohol concentration into the hydrolyzed solution of soysauce koji was 2.6%(v/v) approximately.

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Delivery of Ti Plasmid into Nicotiana sanderae Protoplasts via Liposomes (Liposome을 이용한 Ti Plasmid의 꽃담배 원형질체내 도입)

  • Lim, Myung-Ho;Jeong, Jae-Dong;Kim, In-Soo
    • Applied Biological Chemistry
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    • v.37 no.5
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    • pp.343-348
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    • 1994
  • Ti plasmid of A. tumefaciens was labeled with $^3H-thymidine$, purified and encapsulated into phosphatidylserine (PS) and PS-cholesterol (Chol; 1 : 1 molar ratio) liposomes by lyophilization-rehydration method. PS was supplemented with 1 mole percent octadecyl rhodamine B for fluorometric measurement of PS. Liposomes entrapping $^3H-Ti plasmid$ were fused with Nicotiana sanderae protoplasts by treating with 5 mM $CaCl_2$ and 10% PEG. The fusion was evidenced by fluorescence microscopic technique. The amounts of Ti plasmid and PS associated with protoplasts were assayed by the radioactivity of $^3H-Ti plasmid$ and by the fluorescence of rhodamine B. About 7.9% of the PS liposome and 7.2% of PS-Chol liposome were fused with protoplasts. During the fusion process, about 30% of the liposomal contents of PS-Chol liposome was leaked, in contrast to about 60% leakage of its contents in PS liposome. Accounting the number of liposomes fused with protoplasts together with the encapsulation efficiency and the leakage of liposomal contents, it was calculated that ca. 1,700 Ti plasmid was transfered into one protoplast by the present method. This result may indicates that the present method transfers enough Ti plasmid into plant protoplast to elicit genetic transformation of plants.

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Optimization of Encapsulation Conditions for Fermented Red Ginseng Extracts by Using Cyclodextrin (Cyclodextrin을 이용한 발효홍삼농축액 최적 포접 조건)

  • Shin, Myung-Gon;Lee, Gyu-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.11
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    • pp.1708-1714
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    • 2015
  • Fermented red ginseng concentrate is known as a healthy food source, whereas it has off-flavor such as bitterness and sour flavor based on fermentation. ${\beta}$- and ${\gamma}$-cyclodextrin (CD) were used to encapsulate the off-flavor of fermented red ginseng concentrate by using response surface methodology design on ${\beta}$- and ${\gamma}-CD$ combination. The reducing effects were analyzed by sensory evaluation for bitter and sour tastes, ginsenoside Rb1, and total acidity. The optimized mixing ratio of ${\beta}$- and ${\gamma}-CD$ for reducing bitterness was the least expected value of 2.07 at ${\beta}-CD$ 3.74% versus the soluble solid content of fermented red ginseng concentrate and the ${\gamma}-CD$ 20.63% mixture. The encapsulation effects of ginsenoside Rb1 were the most expected value of 96.75% at ${\beta}-CD$ 3.47% and ${\gamma}-CD$ 19.89% mixture. The encapsulation effects of sour taste were the least expected value of 5.63 at ${\beta}-CD$ 9.34% and ${\gamma}-CD$ 9.96% mixture. The encapsulation effects of lactic acid were the most expected value of 67.73% at ${\beta}-CD$ 16.0% and ${\gamma}-CD$ 13.18% mixture. Based on encapsulation and each optimized combination, the most effective entrapping ${\beta}$-and ${\gamma}-CD$ combination ratio was ${\beta}-CD$ 10% and ${\gamma}-CD$ 13%.