• Title/Summary/Keyword: cell immobilization

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A New Approach to Produce Resveratrol by Enzymatic Bioconversion

  • Che, Jinxin;Shi, Junling;Gao, Zhenhong;Zhang, Yan
    • Journal of Microbiology and Biotechnology
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    • v.26 no.8
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    • pp.1348-1357
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    • 2016
  • An enzymatic reaction system was developed and optimized for bioconversion of resveratrol from glucose. Liquid enzyme extracts were prepared from Alternaria sp. MG1, an endophytic fungus from grape, and used directly or after immobilization with sodium alginate. When the enzyme solution was used, efficient production of resveratrol was found within 120 min in a manner that was pH-, reaction time-, enzyme amount-, substrate type-, and substrate concentration-dependent. After the optimization experiments using the response surface methodology, the highest value of resveratrol production (224.40 μg/l) was found under the conditions of pH 6.84, 0.35 g/l glucose, 0.02 mg/l coenzyme A, and 0.02 mg/l ATP. Immobilized enzyme extracts could keep high production of resveratrol during recycling use for two to five times. The developed system indicated a potential approach to resveratrol biosynthesis independent of plants and fungal cell growth, and provided a possible way to produce resveratrol within 2 h, the shortest period needed for biosynthesis of resveratrol so far.

Optimal Conditions of Co-Immobilized Mixed Culture System with Aspergillus awamori and Zymomonas mobilis (Aspergillus awamori와 Zymomonas mobilis로 구성된 혼합고정화 배양계의 최적 조건)

  • 박석규;이상원;손봉수;최수철;서권일;성낙계;김홍출
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.5
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    • pp.803-810
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    • 1995
  • Co-immobilized mixed culture system(A-Z system) composed of two different oxygen-demanding strains, aerobic(Aspergillus awamori) and anaerobic(Zymomonas mobilis) strains, in a Ca-alginate gel beads was developed to increase ethanol production from raw starch as a carbon source. Optimal mixture ratio of A. awamori and Z. mobilis was $1.25{\times}10^{9}\;spores/L-gel$ and 0.5g cells/L-gel, respectively. After 120 hours of cultivation, gel beads distinguished oxygen-rich surface for A. awamori from oxygen-deficient central part for Z. mobilis. At A-Z culture system, yield of ethanol on glucose, $Y_{p/s}=0.18$, was very low and there was high leakage of cells from surface of gel beads. At A-Z 36 cultrue system with changing silicon check valve for cotton plug at 36 hours in A-Z culture system, there was no cell leakage from gel beads, pH was maintained at around 4.3 during cultivation, and yield of ethanol on glucose, $Y_{p/s}=0.36$, showed 2 times higher than that of control culture system(cotton plug culture).

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Rapid Detection of Salmonella spp. by Antibody-Immobilized Piezoelectric Crystal Biosensor (고정화법을 달리하여 제조한 압전류적 항체 센서에 의한 Salmonella spp.의 신속 검출)

  • 박인선;김우연;김남수
    • Journal of Food Hygiene and Safety
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    • v.13 no.3
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    • pp.206-212
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    • 1998
  • An improved antibody-coated sensor system based on quartz crystal microbalance was developed for the detection of Salmonella spp. An antibody against Salmonella common structural antigen was immobilized onto one gold electrode of the piezoelectric quartz crystal surface by various immobilization procedures. The best results in sensitivity and stability were obtained with the thin layers of protein A and 3,3'-dithiopropionimidate.2HCI(DTBP), a homobifunctional thiol-cleavable crosslinker. After the addition of a S. typhimurium suspension into a reaction cell with 0.1 M sodium phosphate buffer, pH 7.2, the resonant frequency owing to S. typhimurium adsorption decreased conspicuously. The antibody-immobilized crystals prepared by the gold-protein A complex formation and DTBP thiolation showed the frequency shifts of 80 and 283 Hz, respectively. The time required for maximum frequency shift was about 30~60 min. The antibody-coated crystal could be reused for 6~8 consecutive assays.

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Enhancing the Physical Properties and Lifespan of Bacterial Quorum Quenching Media through Combination of Ionic Cross-Linking and Dehydration

  • Lee, Sang Hyun;Lee, Seonki;Lee, Kibaek;Nahm, Chang Hyun;Jo, Sung-Jun;Lee, Jaewoo;Choo, Kwang-Ho;Lee, Jung-Kee;Lee, Chung-Hak;Park, Pyung-Kyu
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.552-560
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    • 2017
  • Quorum quenching (QQ) bacteria entrapped in a polymeric composite hydrogel (QQ medium) have been successfully applied in membrane bioreactors (MBRs) for effective biofouling control. However, in order to bring QQ technology closer to practice, the physical strength and lifetime of QQ media should be improved. In this study, enforcement of physical strength, as well as an extension of the lifetime of a previously reported QQ bacteria entrapping hollow cylinder (QQ-HC), was sought by adding a dehydration procedure following the cross-linking of the polymeric hydrogel by inorganic compounds like $Ca^{2+}$ and boric acid. Such prepared medium demonstrated enhanced physical strength possibly through an increased degree of physical cross-linking. As a result, a longer lifetime of QQ-HCs was confirmed, which led to improved biofouling mitigation performance of QQ-HC in an MBR. Furthermore, QQ-HCs stored under dehydrated condition showed higher QQ activity when the storage time lasted more than 90 days owing to enhanced cell viability. In addition, the dormant QQ activity after the dehydration step could be easily restored through reactivation with real wastewater, and the reduced weight of the dehydrated media is expected to make handling and transportation of QQ media highly convenient and economical in practice.

Production of casein phosphopeptides using Streptococcus faecalis var. liquefaciens cell immobilization (Streptococcus faecalis var. liquefaciens 전세포 고정화법을 이용한 Casein Phosphopeptides 생산)

  • Lee, Ki-Sung;Shin, Jae-Yoon;Jang, Yi-Hyun;Kweon, Dae-Hyuk;Park, Ki-Moon;Jin, Yong-Su
    • KSBB Journal
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    • v.23 no.1
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    • pp.59-64
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    • 2008
  • Optimum conditions for production of casein phosphopeptides (CPP) from sodium casenate by immobilized cell culture of Streptococcus faecalis var. liquefaciens were investigated. Immobilized cells were made by mixing 60% sodium alginate solution with an equal volume of culture broth at the end of exponential phase and subsequently dropping the mixture into $CaCl_{2}$ solution. Optimum conditions for CPP production by the immobilized cells were the same as those ($50^{\circ}C$, pH 7.0, and 10% substrate concentration) by the crude enzyme solution from the supernatant of culture broth. Optimum loading volume of the immobilized cells into a batch reactor was 30% (w/v). Using a continuous reactor loaded by the immobilized cells under the identified optimal conditions, we were able to produce CPP continuously up to 30 days with a maximum CPP conversion efficiency of 20%.

Biodevice Technology (바이오소자 기술)

  • Choi, Jeong-Woo;Lee, Bum-Hwan
    • Korean Chemical Engineering Research
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    • v.44 no.1
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    • pp.1-9
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    • 2006
  • Biodevices composed of biomolecular layer by mimicking the natural functions of cells and the interaction mechanisms of the constituted biomolecules have been developed in various industrial fields such as medical diagnosis, drug screening, electronic device, bioprocess, and environmental pollution detection. To construct biodevices such as bioelectronic devices (biomolecular diode, bio-information storage device and bioelectroluminescence device), protein chip, DNA chip, and cell chip, biomolecules including DNA, protein, and cells have been used. Fusion technology consisting of immobilization technology of biomolecules, micro/nano-scale patterning, detection technology, and MEMs technology has been used to construct the biodevices. Recently, nanotechnology has been applied to construct nano-biodevices. In this paper, the current technology status of biodevice including its fabrication technology and applications is described and the future development direction is proposed.

Organic Solvent-tolerant Lipase from Pseudomonas sp. BCNU 154 (Pseudomonas sp. BCNU 154 유래의 유기용매 내성 리파아제)

  • Choi, Hye Jung;Hwang, Min Jung;Seo, Jeoung-Yoon;Joo, Woo Hong
    • Journal of Life Science
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    • v.23 no.10
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    • pp.1246-1251
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    • 2013
  • An organic solvent-tolerant lipase of Pseudomonas sp. BCNU 154 that was isolated from wastewater in the industrial complex region had optimal activity at $37^{\circ}C$ and pH 8. This crude extracellular lipase from BCNU 154 exhibited maximum stability in toluene, retaining about 6.01 U/ml (117.53%) activity for 2 h. $Ca^{2+}$, $Mg^{2+}$, $NH_4{^+}$, and $Na^+$ ions and triton X-100 activated the enzymes, whereas $Ba^{2+}$, $Hg^{2+}$, and $Zn^{2+}$ ions inhibited their activity. Pseudomonas sp. BCNU 154 lipase revealed stable activity comparable to that of the commercial immobilized Novozym 435. Thus, this organic solvent-tolerant lipase could have potential as a whole cell biocatalyst in industrial chemical processes without the use of immobilization.

Production, Immobilization, and Characterization of Croceibacter atlanticus Lipase Isolated from the Antarctic Ross Sea (남극 로스해에서 분리한 Croceibacter atlanticus균 유래 리파아제의 생산, 고정화, 효소특성 연구)

  • Park, Chae Gyeong;Kim, Hyung Kwoun
    • Microbiology and Biotechnology Letters
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    • v.46 no.3
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    • pp.234-243
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    • 2018
  • The Antarctic Ocean contains numerous microorganisms that produce novel biocatalysts that can have applications in various industries. We screened various psychrophilic bacterial strains isolated from the Ross Sea and found that a Croceibacter atlanticus strain (Stock No. 40-F12) showed high lipolytic activity on a tributyrin plate. We isolated the corresponding lipase gene (lipCA) by shotgun cloning and expressed the LipCA enzyme in Escherichia coli cells. Homology modeling of LipCA was carried out using the Spain Arreo lake metagenome alpha/beta hydrolase as a template. According to the model, LipCA has an ${\alpha}/{\beta}$ hydrolase fold, Gly-X-Ser-X-Glymotif, and lid sequence, indicating that LipCA is a typical lipase enzyme. Active LipCA enzyme was purified fromthe cell-free extract by ammonium sulfate precipitation and gel filtration chromatography. We determined its enzymatic properties including optimum temperature and pH, stability, substrate specificity, and organic solvent stability. LipCA was immobilized by the cross-linked enzyme aggregate (CLEA) method and its enzymatic properties were compared to those of free LipCA. After cross-linking, temperature, pH, and organic solvent stability increased considerably, whereas substrate specificities did not changed. The LipCA CLEA was recovered by centrifugation and showed approximately 40% activity after 4th recovery. This is the first report of the expression, characterization, and immobilization of a C. atlanticus lipase, and this lipase could have potential industrial application.

Immobilization of Bacillus sp. Strains, Catalase Producing Bacteria and Their Hydrogen Peroxide Removal Characteristics (카탈라제를 생산하는 고초균 (Bacillus sp.)의 고정화 및 과산화수소 분해 특성)

  • Han, Kyung-Ah;Jang, Yun-Hee;Rhee, Jong-Il
    • KSBB Journal
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    • v.25 no.6
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    • pp.520-526
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    • 2010
  • In this work we have investigated the production of catalase from Bacillus sp. strains, which were screened and identified from soil. These strains were cultivated in shaking flasks with tryptic soy broth (TSB) at $30^{\circ}C$ and 200 rpm. Effects of the temperature and pH on the stability of the native catalase and whole cell viability were studied in the temperature range of $25-60^{\circ}C$ and the pH range of 7-13. Korean natural zeolite was added to culture medium and mixed with microorganisms for 24 hours. The native catalase maintained its activity over $50^{\circ}C$. The enzyme acitiviy of the catalase from Bacillus flexus BKBChE-3 was highest among the Bacillus sp. strains studied. Bacillus flexus BKBChE-3 and immobilized Bacillus cells have survived under extreme conditions of over $50^{\circ}C$ and pH 12. 60 mL of 10.5 mM $H_2O_2$ solution were entirely removed within 1 hour with catalase produced from Bacillus sp. on the flask. When Bacillus cells were immobilized on Korean natural zeolite, colony forming unit of Bacillus flexus BKBChE-3 was increased and high efficiency of hydrogen peroxide removal was observed.

Biosorption Characteristics of Pb and Cu by Ca-alginate Immobilized Algae Spirulina platensis (Ca-alginate에 고정한 Spirulina platensis의 납과 구리 흡착 특성)

  • Shin, Taek-Soo;Woo, Byoung-Sung;Lim, Byung-Seo;Kim, Kwang-Yul
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.4
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    • pp.446-452
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    • 2008
  • This study was conducted to research the biosorption characteristics using algae, Spirulina platensis, for the removal of Pb and Cu ions in wastewater. Both of free algal cell and immobilized algae by Ca-alginate were used as bioadsorbent, and experiment was proceed in batch reactor for Pb and Cu ions removal, respectively. In the biosorption of Pb and Cu ions by free Spirulina platensis cell, the adsorption equilibrium reached within 20 minute. The higher adsorbed amount of Pb and Cu was shown as increasing of initial concentration of Pb and Cu, and pH of solution, respectively, and the optimum pH was 4.5$\sim$5.0. Under the conditions of initial concentration of Pb or Cu are 200 mg/L, the maximum amounts of Pb and Cu adsorbed to the unit weight of Spirulina platensis were 86.43 and 57.02 mg/g, respectively, and these values were 1.94 and 1.48 times higher than those of activated carbon under same conditions, respectively. The biosorption kinetics of Pb and Cu ions by free Spirulina platensis cell fitted very well to the Freundlich and Langmuir isotherm. The maximum amount of Pb or Cu adsorbed to the unit mass of adsorbent by the Langmuir isotherm($q_{max}$) represented as 95.24 and 62.50 mg/g, respectively. The FT-IR results of free Spirulina platensis biomass showed that biomass has different functional groups and these functional groups are able to react with metal ions in aqueous solution. In the biosorption of Pb and Cu ions by Ca-alginate immobilized algae Spirulina platensis, the adsorption equilibrium reached within 40 min. and observed a little diffusion limitation differed from the free algal cell adsorption.