• Title/Summary/Keyword: whole cell conversion

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Studies on Microbial Penicillin Amidase (II) Characteristics and the Reactor Performance of Whole Cell Immobilized Penicillin Amidase of Escherichia coli (미생물 페니실린 아미다제에 관한 연구 (II) E. coli의 균체 고정화 페니실린 아미다제의 특성 및 반응조에 관한 연구)

  • Seong, Baik-Lin;Kim, Bong-Hee;Mheen, Tae-Iek;Moon H. Han
    • Microbiology and Biotechnology Letters
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    • v.9 no.1
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    • pp.35-44
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    • 1981
  • Whole cell penicillin amidase of Escherichia coli was immobilized by entrapment in gelatin followed by extrusion and crosslinking with glutaraldehyde. The immobilized engyme preparation demonstrated the recovery yield of activity up to 70% and good stability during storage and operation. The half life of activity decay during the operation was estimated to be about 50 days. The optimum pH and temperature for both of immobilized and soluble enzyme are 8.5 and 5$0^{\circ}C$, respectively. No significant change was demonstrated in the effect of pH and temperature, but the increase in heat stability at high temperature was observed in the case of the immobilized enzyme. It was found that the plug flow reactor could be operated favorably since the pH drop along the column path due to tile reaction product was minimized by employing substrate solution with moderate buffer strength. The optimal condition of reactor operation was discussed with regard to the effect of substrate concentration and the residence time on the conversion efficiency and productivity.

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Functional Study of Lysine Decarboxylases from Klebsiella pneumoniae in Escherichia coli and Application of Whole Cell Bioconversion for Cadaverine Production

  • Kim, Jung-Ho;Kim, Hyun Joong;Kim, Yong Hyun;Jeon, Jong Min;Song, Hun Suk;Kim, Junyoung;No, So-Young;Shin, Ji-Hyun;Choi, Kwon-Young;Park, Kyung Moon;Yang, Yung-Hun
    • Journal of Microbiology and Biotechnology
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    • v.26 no.9
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    • pp.1586-1592
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    • 2016
  • Klebsiella pneumoniae is a gram-negative, non-motile, rod-shaped, and encapsulated bacterium in the normal flora of the intestines, mouth, skin, and food, and has decarboxylation activity, which results in generation of diamines (cadaverine, agmatine, and putrescine). However, there is no specific information on the exact mechanism of decarboxylation in K. pnuemoniae. Specifically lysine decarboxylases that generate cadaverine with a wide range of applications has not been shown. Therefore, we performed a functional study of lysine decarboxylases. Enzymatic characteristics such as optimal pH, temperature, and substrates were examined by overexpressing and purifying CadA and LdcC. CadA and LdcC from K. pneumoniae had a preference for L-lysine, and an optimal reaction temperature of 37℃ and an optimal pH of 7. Although the activity of purified CadA from K. pneumoniae was lower than that of CadA from E. coli, the activity of K. pneumoniae CadA in whole cell bioconversion was comparable to that of E. coli CadA, resulting in 90% lysine conversion to cadaverine with pyridoxal 5'-phosphate L-lysine.

Production of ʟ-Theanine Using Escherichia coli Whole-Cell Overexpressing γ-Glutamylmethylamide Synthetase with Baker's Yeast

  • Yang, Soo-Yeon;Han, Yeong-Hoon;Park, Ye-Lim;Park, Jun-Young;No, So-young;Jeong, Daham;Park, Saerom;Park, Hyung Yeon;Kim, Wooseong;Seo, Seung-Oh;Yang, Yung-Hun
    • Journal of Microbiology and Biotechnology
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    • v.30 no.5
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    • pp.785-792
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    • 2020
  • ʟ-Theanine, found in green tea leaves has been shown to positively affect immunity and relaxation in humans. There have been many attempts to produce ʟ-theanine through enzymatic synthesis to overcome the limitations of traditional methods. Among the many genes coding for enzymes in the ʟ-theanine biosynthesis, glutamylmethylamide synthetase (GMAS) exhibits the greatest possibility of producing large amounts of production. Thus, GMAS from Methylovorus mays No. 9 was overexpressed in several strains including vectors with different copy numbers. BW25113(DE3) cells containing the pET24ma::gmas was selected for strains. The optimal temperature, pH, and metal ion concentration were 50℃, 7, and 5 mM MnCl2, respectively. Additionally, ATP was found to be an important factor for producing high concentration of ʟ-theanine so several strains were tested during the reaction for ATP regeneration. Baker's yeast was found to decrease the demand for ATP most effectively. Addition of potassium phosphate source was demonstrated by producing 4-fold higher ʟ-theanine. To enhance the conversion yield, GMAS was additionally overexpressed in the system. A maximum of 198 mM ʟ-theanine was produced with 16.5 mmol/l/h productivity. The whole-cell reaction involving GMAS has greatest potential for scale-up production of ʟ-theanine.

Effect of the Pore Structure on the Anodic Property of SOFC (SOFC 음극의 기공구조가 음극특성에 미치는 영향)

  • 허장원;이동석;이종호;김재동;김주선;이해원;문주호
    • Journal of the Korean Ceramic Society
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    • v.39 no.1
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    • pp.86-91
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    • 2002
  • Solid Oxide Fuel Cells (SOFC) are of great interest of next generation energy conversion system due to their high energy efficiency and environmental friendliness. The basic SOFC unit consists of anode, cathode and solid electrolyte. Among these components, anode plays the most important role for the oxidation of fuel to generate electricity and also behaves as a substrate of the whole cell. It is normally requested that the anode materials should have the high electrical conductivity and gas permeability to reduce the polarization loss of the cell. In this study, the effect of pore former on the microstructure of anode substrate was investigated and thus on the electrical conductivity and the gas permeability. According to the results, microstructure and electrical conductivity of anode substrate were greatly influenced by the shape of pore former and especially by the anisotrpy of the pore former. The use of anisotropic pore former is supposed to deteriorate the cell performance by which the electrical conduction path is disconnected but also the effective gas diffusion path for the fuel is reduced.

Continuous Fermentationof L-Lysine by Immobilized Corynebacterium glutamicum (Corynebacterium glutamicum 고정화균체에 의한 L-라이신 연속발효)

  • 이인선;조정일
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.23 no.2
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    • pp.322-327
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    • 1994
  • For the improvement of L-lysine productivity, development of the continuous fermentation system by a bioreactor assembly was attempted. Primarily, optimal conditions on the whole cell immobilization of Corynebacterium glutamicum ATCC21514 were studied and 76.2% of immobilization ratio was obtained when the cells were entrapped with 4% k-carrageenan showing 4.0kg gel strength. A bioreactor system was set up using the immobilized cells was applied for the continuous production of L-lysine. The results obtained under the optimum conditions were compared with those of the batchwise fermentation. Experimental results obtained from 14 day continuous fermentation showed 36.7% of sugar conversion to L-lysine while the productivity of L-lysine was disclosed as 4.96mg/ml mg-dry cell weight /hr which is 2.5times and 4.1 times higher than those of the batchwise fermentation by the intact cells and by the immobilized cells, respectively.

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Characterizations of i-a-Si:H and p-a-SiC:H Film using ICP-CVD Method to the Fabrication of Large-area Heterojunction Silicon Solar Cells

  • Jeong, Chae-Hwan;Jeon, Min-Sung;Kamisako, Koichi
    • Transactions on Electrical and Electronic Materials
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    • v.9 no.2
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    • pp.73-78
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    • 2008
  • We investigated for comparison of large-area i-a-Si:H and p-a-SiC:H film quality like thickness uniformity, optical bandgap and surface roughness using both ICP-CVD and PECVD on the large-area substrate(diameter of 100 mm). As a whole, films using ICP-CVD could be achieved much uniform thickness and bandgap of that using PECVD. For i-a-Si:H films, its uniformity of thickness and optical bandgap were 2.8 % and 0.38 %, respectively. Also, thickness and optical bandgap of p-a-SiC:H films using ICP-CVD could be obtained at 1.8 % and 0.3 %, respectively. In case of surface roughness, average surface roughness (below 5 nm) of ICP-CVD film could be much better than that (below 30 nm) of PECVD film. HIT solar cell with 2 wt%-AZO/p-a-SiC:H/i-a-Si:H/c-Si/Ag structure was fabricated and characterized with diameter of 152.3 mm in this large-area ICP-CVD system. Conversion efficiency of 9.123 % was achieved with a practical area of $100\;mm\;{\times}\;100\;mm$, which can show the potential to fabrication of the large-area solar cell using ICP-CVD method.

An Approach for Lactulose Production Using the CotX-Mediated Spore-Displayed β-Galactosidase as a Biocatalyst

  • Wang, He;Yang, Ruijin;Hua, Xiao;Zhang, Wenbin;Zhao, Wei
    • Journal of Microbiology and Biotechnology
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    • v.26 no.7
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    • pp.1267-1277
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    • 2016
  • Currently, enzymatic synthesis of lactulose, a synthetic prebiotic disaccharide, is commonly performed with glycosyl hydrolases. In this work, a new type of lactulose-producing biocatalyst was developed by displaying β-galactosidase from Bacillus stearothermophilus IAM11001 (Bs-β-Gal) on the surface of Bacillus subtilis 168 spores. Localization of β-Gal on the spore surface as a fusion to CotX was verified by western blot analysis, immunofluorescence microscopy, and flow cytometry. The optimum pH and temperature for the resulting spore-displayed β-Gal was 6.0 and 75℃, respectively. Under optimal conditions, it showed maximum activity of 0.42 U/mg spores (dry weight). Moreover, the spore-displayed CotX-β-Gal was employed as a whole cell biocatalyst to produce lactulose, yielding 8.8 g/l from 200 g/l lactose and 100 g/l fructose. Reusability tests showed that the spore-displayed CotX-β-Gal retained around 30.3% of its initial activity after eight successive conversion cycles. These results suggest that the CotX-mediated spore-displayed β-Gal may provide a promising strategy for lactulose production.

Whole-cell Biotransformation of Chlorella Oil Hydrolysates into Medium Chain Fatty Acids

  • Seo, Joo-Hyun;Min, Won-Ki;Lee, Jung-Hoo;Lee, Sun-Mee;Lee, Choul-Gyun;Park, Jin-Byung
    • Journal of Marine Bioscience and Biotechnology
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    • v.10 no.2
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    • pp.44-52
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    • 2018
  • A synthetic pathway, which consisted of fatty acid double bond hydratase, alcohol dehydrogenase, and Baeyer-Villiger monooxygenase, was applied to Chlorella oil to produce ester fatty acids, which can be hydrolyzed into medium chain fatty acids. Since linoleic acid is a major fatty acid constituent of Chlorella oil, a fatty acid double bond hydratase from Lactobacillus acidophilus NBRC13951, which is able to convert linoleic acid into 13-hydroxyoctadec-9-enoic acid, was used. Recombinant Escherichia coli expressing the fatty acid double bond hydratase from L. acidophilus NBRC13951 successfully transformed linoleic acid in Chlorella oil hydrolysates into 13-hydroxyoctadec-9-enoic acid with approximately 60% conversion yield. 13-Hydroxyoctadec-9-enoic acid was further converted into ester fatty acids by the recombinant E. coli expressing a long chain secondary alcohol dehydrogenase and a Baeyer-Villiger monooxygenase. The resulting ester fatty acids were then hydrolyzed into medium chain fatty acids by a lipase. Overall, industrially relevant medium chain fatty acids were produced from Chlorella oil hydrolysates. Thereby, this study may contribute to biosynthesis of medium chain fatty acids from microalgae oils as well as long chain fatty acids.

A Comparative Study of Two Different SnO2:F-coated Glass Substrates for CdTe Solar Cells

  • Cha, Eun Seok;Ko, Young Min;Choi, Yong Woo;Park, Gyu Chan;Ahn, Byung Tae
    • Current Photovoltaic Research
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    • v.5 no.1
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    • pp.1-8
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    • 2017
  • Two different fluorine-doped tin oxide (FTO)-coated glass substrates were investigated to find better suitability for CdTe solar cells. Substrate A consisted of FTO (300 nm)/$SiO_2$ (24 nm)/intrinsic $SnO_2$ (30 nm)/borosilicate glass (2.2 mm), and substrate B consisted of FTO (700 nm)/intrinsic $SnO_2$ (30nm)/borosilicate glass (1.8 mm). The overall thickness of the FTO/glass substrates was about 2.5 mm. The total light transmittance of substrate B was much higher than that of substrate A throughout the whole spectral region, even though the thickness of the FTO in substrate B was twice larger than that of the FTO in the substrate A. The short-circuit current greatly increased in substrate B and the external quantum efficiency (EQE) increased over the whole wavelength range. This study shows that the diffuse optical transmittance played a key role in the large EQE value in the blue wavelength region, and the direct transmittance played a key role in the large EQE value in the red wavelength region. The higher transmittance is due to the rough surface generated by the thicker FTO on glass. The conversion efficiency of the CdTe solar cell increased from 12.4 to 15.1% in combination of rough FTO substrate and Cu solution back contact.

High Purity Hydrogen Generator for Fuel Cell Vehicles (연료전지 자동차 탑재형 고순도 수소생산장치)

  • Han, Jaesung;Lee, Seok-Min
    • Transactions of the Korean hydrogen and new energy society
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    • v.12 no.4
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    • pp.277-285
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    • 2001
  • We developed a compact, 10 kWe, purifier-integrated reformer which supplies hydrogen for fuel cell vehicles. Our proprietary technologies regarding hydrogen purification by palladium alloy membrane and catalytic combustion by noble metal coated wire-mesh catalyst were combined with the conventional methanol steam reforming technology, resulting in higher conversion, excellent quality of product hydrogen, and better thermal efficiency than any other systems. In this system, steam reforming, hydrogen purification, and catalytic combustion take place all in a single reactor so that the whole system is compact and easy to operate. The module produces $8.2Nm^3/hr$ of 99.999% or higher purity hydrogen with CO impurity less than 10 ppm, which is equivalent to 10 kWe when PEMFC has 45 % efficiency. Thermal efficiency of the module is 81 % and the power density of the module is 1.6 L/kWe. As the results of experiments, cold-start time has been measured about 20 minutes. Response time of hydrogen production to the change of the feed rate has been within 1 minutes.

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