• Title/Summary/Keyword: Arabinose

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Microbial production of carotenoids for fortification of foods

  • Kim, Seon-Won;Keasling, J.D.
    • Proceedings of the Korean Society of Life Science Conference
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    • 2001.11a
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    • pp.3-8
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    • 2001
  • Isopentenyl diphosphate (IPP) is the common, five-carbon building block in the biosynthesis of all carotenoids, IPP in Escherichia coli is synthesized through the non-mevalonate pathway. The first reaction of IPP biosynthesis in E. coli is the formation of 1-deoxy-D-xylulose-5-phosphate (DXP), catalyzed by DXP synthase and encoded by dxs. The second reaction in the pathway is the reduction of DXP to 2-C-methyl-D-erythritol-4-phosphate, catalyzed by DXP reductoisomerase and encoded by dxr. To determine if one or more of the reactions in the non-mevalonate pathway controlled flux to IPP, dxs and dxr were placed on several expression vectors under the control of three different promoters and transformed into three E. coli strains (DH5(, XL1-Blue, and JM101) that had been engineered to produce lycopene. Lycopene production was improved significantly in strains transformed with the dxs expression vectors. When the dxs gene was expressed from the arabinose-inducible araBAD promoter (PBAD) on a medium-copy plasmid, lycopene production was 2-fold higher than when dxs was expressed from the IPTG-inducible trc and lac promoters (Ptrc and Plac, respectively) on medium-copy and high-copy plasmids, Given the low final densities of cells expressing dxs from IPTG-inducible promoters, the low lycopene production was probably due to the metabolic burden of plasmid maintenance and an excessive drain of central metabolic intermediates. At arabinose concentrations between 0 and 1.33 mM, cells expressing both dxs and dxr from PBAD on a medium-copy plasmid produced 1.4 - 2.0 times more lycopene than cells expressing dxs only. However, at higher arabinose concentrations lycopene production in cells expressing both dxs and dxr was lower than in cells expressing dxs only. A comparison of the three E. coli strains transformed with the arabinose-inducible dxs on a medium-copy plamid revealed that lycopene production was highest in XL1-Blue.

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Protection of the brain through supplementation with larch arabinogalactan in a rat model of vascular dementia

  • Lim, Sun Ha;Lee, Jongwon
    • Nutrition Research and Practice
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    • v.11 no.5
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    • pp.381-387
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    • 2017
  • BACKGROUND/OBJECTIVES: Vascular dementia (VaD) caused by reduced blood supply to the brain manifests as white matter lesions accompanying demyelination and glial activation. We previously showed that arabinoxylan consisting of arabinose and xylose, and arabinose itself attenuated white matter injury in a rat model of VaD. Here, we investigated whether larch arabinogalactan (LAG) consisting of arabinose and galactose could also reduce white matter injury. MATERIALS/METHODS: We used a rat model of bilateral common carotid artery occlusion (BCCAO), in which the bilateral common carotid arteries were exposed and ligated permanently with silk sutures. The rats were fed a modified AIN-93G diet supplemented with LAG (100 mg/kg/day) for 5 days before and 4 weeks after being subjected to BCCAO. Four weeks after BCCAO, the pupillary light reflex (PLR) was measured to assess functional consequences of injury in the corpus callosum (cc). Additionally, Luxol fast blue staining and immunohistochemical staining were conducted to assess white matter injury, and astrocytic and microglial activation, respectively. RESULTS: We showed that white matter injury in the the cc and optic tract (opt) was attenuated in rats fed diet supplemented with LAG. Functional consequences of injury reduction in the opt manifested as improved PLR. Overall, these findings indicate that LAG intake protects against white matter injury through inhibition of glial activation. CONCLUSIONS: The results of this study support our hypothesis that cell wall polysaccharides consisting of arabinose are effective at protecting white matter injury, regardless of their origin. Moreover, LAG has the potential for development as a functional food to prevent vascular dementia.

Modulation of the Tendency Towards Inclusion Body Formation of Recombinant Protein by the Addition of Glucose in the araBAD Promoter System of Escherichia coli

  • Lee, You-Jin;Jung, Kyung-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1898-1903
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    • 2007
  • We attempted to modulate the overall protein expression rate through the addition of a repressor against the araBAD promoter system of Escherichia coli, in which glucose was used as a repressor. Therefore, 0.5% L-arabinose was initially contained as an inducer in culture medium, and either 2% glucose or 2% glycerol was used as a carbon source, and it was found that the expression of recombinant interferon-${\alpha}$ could be observed at the beginning of the batch culture when glycerol was used as a carbon source. However, when glucose was used, the initiation of recombinant interferon-${\alpha}$ expression was delayed compared with that when glycerol was used. Furthermore, when the addition of 0.5% glucose was carried out once or twice after 0.5% L-arabinose induction during DO-stat fed-batch culture, the distributions of soluble and insoluble recombinant interferon-${\alpha}$ were modulated. When glucose was not added after the induction of L-arabinose, all of the expressed recombinant interferon-${\alpha}$ formed an inclusion body during the later half of culturing. However, when glucose was added after induction, the expressed recombinant interferon-${\alpha}$ did not all form an inclusion body, and about half of the total recombinant interferon-${\alpha}$ was expressed in a soluble form. It was deduced that the addition of glucose after the induction of L-arabinose might lower the cAMP level, and thus, CAP (catabolite activator protein) might not be activated. The transcription rate of recombinant interferon-${\alpha}$ in the araBAD promoter system might be delayed by the partial repression. This inhibition of the transcription rate probably resulted in more soluble interferon-${\alpha}$ expression caused by the reduction of the protein synthesis rate.

Metabolic Engineering of Nonmevalonate Pathway in Escherichia coli Enhances Lycopene Production

  • Kim, Seon-Won;J.D. Keasling
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2001.06a
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    • pp.141-145
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    • 2001
  • Isopentenyl diphosphate (IPP) is the common, five-carbon building block in the biosynthesis of all carotenoids. IPP in Escherichia coli is synthesized through the non-mevalonate pathway. The first reaction of IPP biosynthesis in E. coli is the formation of l-deoxy-D-xylulose-5-phosphate (DXP), catalyzed by DXP synthase and encoded by dxs. The second reaction in the pathway is the reduction of DXP to 2-C-methyl-D-erythritol-4-phosphate, catalyzed by DXP reductoisomerase and encoded by dxr. To determine if one or more of the reactions in the non-mevalonate pathway controlled flux to IPP, dxs and dxr were placed on several expression vectors under the control of three different promoters and transformed into three E. coli strains (DH5$\alpha$, XL1-Blue, and JMl0l) that had been engineered to produce lycopene. Lycopene production was improved significantly in strains transformed with the dxs expression vectors. When the dxs gene was expressed from the arabinose-inducible araBAD promoter ( $P_{BAD}$) on a medium-copy plasmid, lycopene production was 2-fold higher than when dxs was expressed from the IPTG-inducible trc and lac promoters ( $P_{trc}$ and $P_{lac}$, respectively) on medium-copy and high-copy plasmids. Given the low final densities of cells expressing dxs from IPTG-inducible promoters, the low lycopene production was probably due to the metabolic burden of plasmid maintenance and an excessive drain of central metabolic intermediates. At arabinose concentrations between 0 and 1.33 roM, cells expressing both dxs and dxr from $P_{BAD}$ on a medium-copy plasmid produced 1.4 - 2.0 times more lycopene than cells expressing dxs only. However, at higher arabinose concentrations lycopene . production in cells expressing both dxs and dxr was lower than in cells expressing dxs only. A comparison of the three E. coli strains transformed with the arabinose-inducible dxs on a medium-copy plasmid revealed that lycopene production was highest in XLI-Blue.LI-Blue.

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Isolation and Characterization of Oil Degrading Bacteria from Southern Sea of Korea (남해안 해수로부터 원유 분해 세균의 분리 및 특성)

  • 김학주;김봉조;공재열;구헌서
    • KSBB Journal
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    • v.15 no.1
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    • pp.27-34
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    • 2000
  • A marine bacterium having a high oil-degrading activity was isolated form the oil-polluted southern sea of Korea, and was identified as Pseudomonas aeruginosa and was named Pseudomonas aeruginosa BYK-2. The optimal tmeperatur, culture time, pH and NaCl concentration for biosurfactant production and cell growth showed $25^{\circ}C$, 48h, 7.0 and 0%(w/v), respectively. After cultivation at $25^{\circ}C$, 180 rpm in 250 mL erlenmeyer flask for 7days, 1%(w/v) arabian light crude oil and bunker C oil which are considered to be hardly degradable compounds were degraded 92.1%(w/w) and 76%(w/w) respectively. And then, cell adherence was measured on various carbon sources. The cell adherence indicated over 80% on hydrocarbons(arabian light crude oil, kuwait curde oil, bunker C oil, n-paraffine, n-hexadecane, n-tetradecane) as carbon sources. Lecithin among fatty acids(oleic acid, olive oil, lecithin) showed highest cell adherence of 91.5%. The cell adherence of sugars(arabinose, trehalose, dextrose, galactose, lactose, fructose, maltose, sorbitol, sucrose) observed to be less than 70% except for arabinose, galactose, sorbitol and sucrose.

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Metabolic engineering for isoprenoids production in Escherichia coli

  • Kim, Seon-Won;Keasling, J.D.
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.70-73
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    • 2001
  • Isopentenyl diphosphate (IPP) is the common, five-carbon building block in the biosynthesis of all isoprenoids. IPP in Escherichia coli is synthesized through the non-mevalonate pathway. The first reaction of IPP biosynthesis in E. coli is the formation of 1-deoxy-D-xylulose-5-phosphate(DXP), catalyzed by DXP synthase and encoded by dxs. The second reaction in the pathway is the reduction of DXP to 2-C-methyl-D-erythritol-4-phosphate, catalyzed by DXP reductoismerase and encoded by dxr. To determine if one of more of the reactions in the non-mevalonate pathway controlled flux to IPP, dxs and dxr were placed on several expression vectors under the control of three different promoters and transformed into three E. coli strains ($DH5{\alpha}$, XL1-Blue, and JM101) that had been engineered to produce lycopene, a kind of isoprenoids. Lycopene production was improved significantly in strains transformed with the dex expression vectors. At arabinose concentrations between 0 and 1.33 mM, cells expressiong both dxs and from $P_{BAD}$ on a midium-copy plasmid produced 1.4 -2.0 times more lycopene than cells expressing dxs only. However, at higher arabinose concentrations lycopene production in cell expressing both dxs and dxr was lower than in cells expression dxs only. A comparison of the three E. coli strains trasfomed with the arabinose-inducible dxs on a medium-copy plasmid revealed that lycopene production was highest in XL1-Blue.

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The Contents of Free Sugar and Alcohol in Traditional Soy Sauce Prepared from Meju under Different Formations (형상이 다른 메주로 제조한 재래식 간장 중의 유리당과 알코올 함량)

  • 서정숙;이택수
    • The Korean Journal of Food And Nutrition
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    • v.6 no.2
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    • pp.103-108
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    • 1993
  • The Kinds of soy sauce were prepared using the brick type of conventional meju(A), the brick type of meju of Aspergillus oryzae(B), and the grain type of meju of Aspergillus oryzae(C). Free sugar and alcohol were analyzed in accordance to aging time of those products. Galactose, glucose, arabinose, xylose, and mannose were detected in all kinds of soy sauce. In soy sauce 4 galactose, glucose, arabinose, and xylose were shown in the order of content. The content of galactose in soy sauce A and soy sauce C were shown higher than that in soy sauce B alt during the time. The content of glucose was highest among that of all kinds of free sugars at the beginning of preparation, but the content of glucose was lower than that of arabinose and xylose after 60 days. The contents order of total free sugar was soy sauce C > soy sauce B > soy sauce A. The content of ethyl alcohol was shown 6.23∼19.10mg%, and that of isobutyl alcohol was 8.70∼ 169.03 ppm in all soy sauces, but those of soy sauce C was higher than those of other soy sauces.

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Enhancement of Biocontrol Activity of Antagonistic Chryseobacterium Strain KJ1R5 by Adding Carbon Sources against Phytophthora capsici

  • Kim, Yu-Seok;Jang, Bo-Ra;Chung, Ill-Min;Sang, Mee-Kyung;Ku, Han-Mo;Kim, Ki-Deok;Chun, Se-Chul
    • The Plant Pathology Journal
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    • v.24 no.2
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    • pp.164-170
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    • 2008
  • Carbon utilization by Chryseobacterium strain KJ1R5 was studied to enhance its biocontrol activity against Phytophthora capsid. Chryseobacterium strain KJ1R5 has previously been shown to control Phytophthora blight of pepper (Capsicum annuum L.). Strain KJ1R5 could utilize carbon sources such as L-arabinose, D-cellobiose, ${\beta}-lactose$ and D-galactose well. P. capsici could utilize D-glucose well, showing the absorbencies ranged from 0.577 to 0.767 at 600nm. When 2% L-arabinose, which could only be utilized by the bio-control strain KJ1R5, was amended into the bacterial suspension, the efficacy of biological control increased. Among the amendments of various carbon sources into bacterial suspension, L-arabinose and D-(+)-glucose significantly enhanced biological control activity, resulting in a reduction of disease incidence to 6.9%, compared to 21.9% for the strain KJ1R5 alone and 81.3% for P. capsici inoculation alone, indicating that amendment with specific carbon sources could increase the biological control activity.

Physiology of Rhizoctonia solani AG2-2(IV), Trichoderma harzianum, and Chaetomium cochliodes, and their Utilization of Thatch-related Carbohydrate in Zoysia japonica (Rhizoctonia solani AG2-2(IV), Trichoderma harzianum and Chaetomium cochliodes의 생육생리와 이들 미생물들의 한국잔디 대취층 관련 탄소원 이용도 조사)

  • 박진희;강시용;김희규
    • Asian Journal of Turfgrass Science
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    • v.12 no.4
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    • pp.211-220
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    • 1998
  • Cellulose-degrading fungi were idenfied as Rhizoctonia solani AG2-2(IV), T. harzianum and C. cochliodes. Rhizoctonia solani AG2-2(IV) grows better in the acidified media of pH 4 and 5 than pH 6 and 7. Mycelial growth of T. harzianum and C. cochliodes was also higher in pH 4 and 5 than in pH 6 and 7. In order to relate the above findings to nutrient utilization, mycelial growth of R. solani AG2-2(IV) are evaluated with various carbon sources. R. solani AG2-2(IV) grows well in the order of mannose, cellobiose, glucose, xylose and arabinose. However, mycelial dry weights of T. harzianum were 98.7, 78.0, 72.3, 43.7 and 32.3mg in glucose, mannose, cellobiose, xylose, and arabinose, respectively. Mycelial dry weight of C. cochilodes was 118, 65, 57, 49, and 16mg in mannose, cellobiose, xylose, glucose, and arabinose, respectively. Result of cellulase assay of R. solani AG2-2(IV) and soil fungi was reffered as, R. solani AG2-2(IV) produced more cellulase on CMC substrate than on CEL and secretes more enzyme in floated condition than in water-immersed condition. T. harzianum secreted less amount of cellulase than R. solani AG2-2 and C. cochliodes. T. harzianum produced no enzyme on CEL under water-immersed condition. C. cochliodes produced similar amounts of cellulase on either CMC or CEL under both water-immersed and floated condition.

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