• Title/Summary/Keyword: Systems Biotechnology

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Isolation and Characterization of a Novel Agar-Degrading Marine Bacterium, Gayadomonas joobiniege gen, nov, sp. nov., from the Southern Sea, Korea

  • Chi, Won-Jae;Park, Jae-Seon;Kwak, Min-Jung;Kim, Jihyun F.;Chang, Yong-Keun;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.23 no.11
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    • pp.1509-1518
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    • 2013
  • An agar-degrading bacterium, designated as strain $G7^T$, was isolated from a coastal seawater sample from Gaya Island (Gayado in Korean), Republic of Korea. The isolated strain $G7^T$ is gram-negative, rod shaped, aerobic, non-motile, and non-pigmented. A similarity search based on its 16S rRNA gene sequence revealed that it shares 95.5%, 90.6%, and 90.0% similarity with the 16S rRNA gene sequences of Catenovulum agarivorans $YM01^T$, Algicola sagamiensis, and Bowmanella pacifica W3-$3A^T$, respectively. Phylogenetic analyses demonstrated that strain $G7^T$ formed a distinct monophyletic clade closely related to species of the family Alteromonadaceae in the Alteromonas-like Gammaproteobacteria. The G+C content of strain $G7^T$ was 41.12 mol%. The DNA-DNA hybridization value between strain $G7^T$ and the phylogenetically closest strain $YM01^T$ was 19.63%. The genomes of $G7^T$ and $YM01^T$ had an average ANIb value of 70.00%. The predominant isoprenoid quinone of this particular strain was ubiquinone-8, whereas that of C. agarivorans $YM01^T$ was menaquinone-7. The major fatty acids of strain $G7^T$ were Iso-$C_{15:0}$ (41.47%), Anteiso-$C_{15:0}$ (22.99%), and $C_{16:1}{\omega}7c/iso-C_{15:0}2-OH$ (8.85%), which were quite different from those of $YM01^T$. Comparison of the phenotypic characteristics related to carbon utilization, enzyme production, and susceptibility to antibiotics also demonstrated that strain $G7^T$ is distinct from C. agarivorans $YM01^T$. Based on its phenotypic, chemotaxonomic, and phylogenetic distinctiveness, strain $G7^T$ was considered a novel genus and species in the Gammaproteobacteria, for which the name Gayadomonas joobiniege gen. nov. sp. nov. (ATCC BAA-2321 = $DSM25250^T=KCTC23721^T$) is proposed.

Resveratrol Upregulates p21, Cell Cycle Regulator, in Gaucher Disease Cells (Gaucher병에서 resveratrol의 세포주기 조절자 p21을 통한 세포보호 효과 연구)

  • Kim, Dong-Hyun;Heo, Tae-Hwe;Kim, June-Bum;Kim, Sung-Jo
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1281-1286
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    • 2010
  • Gaucher disease (GD) is caused by glucocerebrosidase functional deficiency and the most prevalent lysosomal storage disorder (LSD), with an incidence of about 1 in 20,000 new births. Resveratrol, one kind of phytoalexin, is a produced naturally by several plants and has anti-tumor, anti-aging, anti-inflammatory and neuro-protective effects. In this paper we provide the cellular protective effect of resveratrol in both type I and type II Gaucher disease cells. Resveratrol treatment did not show any significant change in the p21 and p53 mRNA expression level, however expression level of the p21 protein, a cell cycle arrest factor, shows significant increment in both types of Gaucher disease cells. These cell cycle arrest patterns were confirmed by both MTT assay measurement and microscopy detection. In comparison, expression level of poly ADP ribose polymerase (PARP), an apoptosis indicator protein, was significantly decreased in both type I and II Gaucher disease cells after treatment with resveratrol. This result indicates that resveratrol relievescellular apoptotic stress fromtype I and II Gaucher disease cells. Therefore, we demonstrate that resveratrol inhibits cell proliferation via p21 activity and activates cellular repair systems for Gaucher disease cells. Our results provide at least one of the molecular mechanisms of Gaucher disease and may allow the verification of potential drug targets for therapeutic trials.

Industrial Application for Aquaculture of Fermented Soybean with Bacillus polyfermenticus SCD (양식업에서 바실러스 폴리퍼멘티쿠스 콩발효물의 상업적 응용)

  • Lee, Jin Young;Kim, Kang Min;Kang, Jae Seon
    • Journal of Marine Bioscience and Biotechnology
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    • v.2 no.1
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    • pp.48-54
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    • 2007
  • Bacillus polyfermenticus SCD (B. polyfermenticus SCD) has been appropriately used for the therapy of long-term intestinal disorders, because live strains in the form of active endospores can successfully reach the target intestine in humans. B. polyfermenticus SCD produces the most antibiotic-related materials. In the present study, B. polyfermenticus SCD was fermented with soybeans (BPFS) and its probiotic properties were investigated. B. polyfermenticus SCD and BPFS showed a broad spectrum of antimicrobial activity against pathogenic Gram-positive (Streptococcus parauberis, Streptococcus iniae, Lactococcus garviae) and Gram-negative (Flexibacter tractuosus, Vibrio harveyi, Vibrio vulnificus, Vibrio ordalii) bacteria and moulds (Aspergillus niger, Aspergillus oryzae). Sebastes schlegeli were used to examine survival rate and cleanup action by BPFS. Bacterial infection resulted in a mortality of up to 99% in the commercial fodder fed groups. BPFS both enhanced the growth rate of fry by improving their appetite and had cleaned by decreased eutrophication. Therefore, it seems appropriate that BPFS should be developed as an antibiotic replacement, favorable fodder additive, and antifungal material in fish farming systems.

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Exposure and Risk Assessment of Melamine in Representative Korean Foods for Infants and Children (WHO 내용일일섭취량을 고려한 우리나라 영유아 어린이의 식품을 통한 멜라민 노출 및 위해 평가)

  • Oh, Chang-Hwan
    • Journal of Dairy Science and Biotechnology
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    • v.27 no.1
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    • pp.1-12
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    • 2009
  • In 2008, baby formula containing melamine was found to be responsible for a large outbreak of renal failure in infants in China. A total of 294,000 infants were hospitalized, and at least 6 babies died due to ingestion of the tainted formula. Melamine contains high levels of nitrogen (>60%), which is used as an indicator of protein content. Therefore, high levels of melamine in infant formula were thought to be the result of deliberate contamination m an attempt to increase its apparent protein content. Following inspections by China's national inspection agency, assorted products from at least 22 dairy manufacturers across China were found to have varied levels of melamine (range: 0.096196.61 mg/kg). Melamine co-exposure with cyanuric acid can induce acute melamine-cyanurate crystal nephropathy, which can lead to renal failure at much lower doses than if either compound were ingested alone. However, currently, there are very few data on melamine analogues other than cyanuric acid. At an expert meeting of the WHO and FAO held to review toxicological aspects of melamine and cyanuric acid on December 14, 2008, a new tolerable daily intake (TDI) of melamine was established that could be applied to the entire population, including infants. Therefore, a risk assessment of the various theoretical melamine contamination levels in infant formula and selected representative foods (other than infant formula and sole-source nutrition products) is urgently needed for Korean babies and children up to 7 years of age. Although the undetectable level regulation for infant formula may be low enough to guarantee the safety of babies under the age of 1 year (including premature babies), the melamine standard of 2.5 ppm for foods other than baby formula could be insufficient to protect the 95th percentile population aged 1~2 years because of this demographic's high consumption of milk, yogurt, and soy milk (hazard index = 1.79). Because TDIs are chronic values intended to protect an individual over his/her lifetime, occasional modest ingestion in excess of the TDI is not likely to be a health concern. However, children aged 1~2 years may have renal systems that are comparatively more sensitive to the crystallization of melamine and its analogues. Therefore, governmental jurisdictions may need to practice more prudent management of food items that could raise the melamine exposure for this population.

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Water relations of plants under environmental stresses: role of aquaporins

  • Kang, H.S.;Ahn, S.J.;Hong, S.W.;Chung, G.C.
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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    • pp.71-80
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    • 2005
  • Effects of low temperature ($8^{\circ}C$) on the hydraulic conductivity of young roots of a chilling-sensitive (cucumber; Cucumis sativus L.) and a chilling-resistant (figleaf gourd; Cucurbita ficifolia Bouche) crop have been measured at the levels of whole root systems (root hydraulic conductivity, $Lp_r$) and of individual cortical cells (cell hydraulic conductivity, Lp). In figleaf gourd, there was a reduction only in hydrostatic $Lp_r$ but not in osmotic $Lp_r$ suggesting that the activity of water channels was not much affected by low root temperature (LRT)treatment in this species. Changes in cell Lp in response to chilling and recovery were similar asroot level, although they were more intense at the root level. Roots of figleaf gourd recovered better from LRT treatment than those of cucumber. In figleaf gourd, recovery (both at the root and cell level) often resulted in Lp and $Lp_r$ values which were even bigger than the original, i.e. there was an overshoot in hydraulic conductivity. These effects were larger forosmotic (representing the cell-to-cell passage of water) than for hydrostatic $Lp_r$. After a short term (1 d) exposure to $8\;^{\circ}C$ followed by 1 d at $20\;^{\circ}C$, hydrostatic $Lp_r$ of cucumber nearly recovered and that of figleaf gourd still remained higher due to the overshoot. On the contrary, osmotic $Lp_r$ and cell Lp in both species remained high by a factor of 3 as compared to the control, possibly due to an increased activity of water channels. After pre-conditioning of roots at LRT, increased hydraulic conductivitywas completely inhibited by $HgCl_2$ at both the root and cell levels. Different from figleaf gourd, recovery from chilling was not complete in cucumber after longer exposure to LRT. It is concluded that at LRT, both changes in the activity of aquaporins and alterations of root anatomy determine the water uptake in both species. To better understand the aquaporin function in plants under various stress conditions, we examined the transgenic Arabidopsisand tobacco plants that constitutively overexpress ArabidopsisPIP1;4 or PIP2;5 under various abiotic stress conditions. No significant differences in growth rates were found between the transgenic and wild-type plants under favorable growth conditions. By contrast, overexpression of PIP1;4 or PIP2;5 had a negative effect on seed germination and seedling growth under drought stress, whereas it had a positive effect under cold stress and no effect under salt stress. Measurement of water transport by cell pressure probe revealed that these observed phenotypes under different stress conditions were closely correlated with the ability of water transport by each aquaporin in the transgenic plants. Together, our results demonstrate that PIP-type aquaporins play roles in seed germination, seedling growth, and stress response of Arabidopsis and tobacco plants under various stress conditions, and emphasize the importance of a single aquaporin-mediated water transport in these cellular processes.

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In vitro propagation of oil palm (Elaeis guineensis Jacq.) clones through somatic embryogenesis and analysis of somaclonal variation by RAPD (체세포배발생을 통한 오일팜나무(Elaeis guineensis Jacq.) 클론의 기내증식 및 RAPD를 이용한 체세포변이의 검정)

  • Ahn, In-Suk;Park, Hye-Rim;Son, Sung-Ho
    • Journal of Plant Biotechnology
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    • v.39 no.3
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    • pp.196-204
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    • 2012
  • This study was carried out to develop reliable systems for somatic embryogenesis in oil palm tree (Elaeis guineensis Jacq.), and to verify the somaclonal variants by RAPD analysis. Embryogenic callus was induced successfully on modified half-strength MS medium containing $NaH_2PO_4{\cdot}2H_2O$ and casein. Embryogenic callus was further developed to somatic embryo mass (SEM), which is very hard and bonded tightly each other. Plantlets were proliferated when SEM was cultured on modified MS medium containing half strength $NH_4NO_3$, casein and L-ascorbic acid. Plantlets were transplanted into pots containing artificial soils. When RAPD analysis was conducted using randomly selected 95 in vitro plantlets and 19 random primers, somaclonal variation was detected using BNR35 primer. There was missing band around 1 kb in #22, #28, #35, and #77 plantlets. In addition, bands obtained from #28, #35, and #77 was much stronger than other normal bands. The blast results at NCBI revealed that somaclonal variation observed in this study was related to chloroplast genome of oil palm. The results also revealed that oil palm reproduction system through somatic embryogenesis is quite reliable and early detection of somaclonal variants seem to be possible at in vitro stage by RAPD analysis.

Biosynthesis of recombinant human prominiinsulin in E. coli and plant systems (대장균과 식물시스템에서 재조합 인간 prominiinsulin 생합성 분석)

  • Choi, Yu Jin;Park, Su Hyun;Kim, Ji Su;Wi, Soo Jin;Park, Ky Young
    • Journal of Plant Biotechnology
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    • v.40 no.3
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    • pp.169-177
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    • 2013
  • Recently, the number of people with diabetes is rapidly increasing, coupled with the fact that the insulin market is remarkably increasing. Therefore, molecular farming for plant-derived pharmaceutical protein production is reported as becoming more attractive than ever. In this study, we carried out experiments step by step for development of recombinant insulin constructs, which were transformed into E. coli system, in vitro transcription and translation system, and tobacco cells. At first, recombinant proinsulin protein was successfully produced in in vitro transcription and translation system with wheat germ extract. After which, recombinant construct of prominiinsulin encoded a fusion protein of 7.8 kDa with trypsin cleavage sites at N terminus and C terminus of minimized C-peptide was tried to in vitro expression using E.coli culture. After purification with His-tag column, the resulting recombinant prominiinsulin protein was processed with trypsin, and then checked insulin biosynthesis by SDS-PAGE and western blot analysis with anti-insulin monoclonal antibody. The immunoreactive product of trypsin-treated miniinsulin was identical to the predicted insulin hexamer. The construct of 35S promoter-driven preprominiinsulin recombinant gene with signal peptide region for ER-targeting and red fluorescence protein gene [N terminus ${\rightarrow}$ tobacco E2 signal peptide ${\rightarrow}$ B-peptide (1-29 AA) ${\rightarrow}$ AAK ${\rightarrow}$ A-peptide (1-21 AA) ${\rightarrow}$ RR ${\rightarrow}$ His6 ${\rightarrow}$ KDEL ${\rightarrow}$ C terminus] was transformed into BY-2 tobacco cells. A polypeptide corresponding to the 38-kDa molecular mass predicted for fusion protein was detected in total protein profiles from transgenic BY-2 cells by western analysis. Therefore, this recombinant preprominiinsulin construct can be used for generation of transgenic tobacco plants producing therapeutic recombinant insulin.

Research Priorities to Support Mandatory Implementation of a Total Pollutant Load Management System (TPLMS) in the Han River Basin (한강수계 의무적 수질오염총량관리제시행지원을 위한 조사·연구의 우선순위 설정)

  • Lee, Chang-Hee;Lee, Bum-Yeon;Lee, Su-Woong
    • Journal of Environmental Policy
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    • v.8 no.4
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    • pp.25-36
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    • 2009
  • The Total Pollutant Load Management System(TPLMS) in the Han River basin is being changed from a voluntary to a mandatory system. Accordingly, this study suggests directions and priorities for research that can support implementation of TPLMS through an objective approach that deploys gap analysis and analytic hierarchy processes (AHP). Gap analysis indicated that TPLMS in Korea is still focused on compliance with regulations, and that implementation of TPLMS is still in its early stage. Improvements are thus needed in flexibility and effectiveness, including introduction of emissions rights trading, and upgrading to a renewable emissions permit system. The AHP study indicated that R&D will need to proceed in parallel in multiple areas to improve systems and resolve scientific uncertainties. Balanced R&D will be needed in both the institutional and technical groups. Subgroup analysis indicated that developing a reasonable process to establish water quality management targets is of the highest priority in the institutional group. In the technical group, higher priority will need to be given to improving model reliability and developing innovative pollution load reduction technologies.

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Effects of a Herbal Composition (HemoHIM) on the Activation of Dendritic Cells (생약복합조성물(HemoHIM)의 수지상세포 활성화 효과)

  • Shin, Sung-Hae;Kim, Do-Soon;Kim, Sung-Ho;Jo, Sung-Kee;Byun, Mung-Woo;Yee, Sung-Tae
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.10
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    • pp.1322-1328
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    • 2006
  • In our previous study, a novel herb mixture (HIM-I) of Angelica gigas radix, Cnidium officinale rhizoma, and Paeonia japonica radix was developed to protect= the intestinal and immune systems and to promote their recovery from radiation damage. A new herbal composition (HemoHIM) with the high immune modulating activity was developed from HIM-I. In the present study, we examined the effects of HemoHIM on the maturation process of murine bone marrow (BM)-derived dendritic cells (DC). BM cells were cultured in the presence of iL-4 and GM-CSF and the generated immature DC were stimulated with HemoHIM for 24 hours. HemoHIM significantly enhanced the expression of co-stimulatory molecules, CD80 and CD86, especially. The activation capacity of HemoHIM-treated DC was significantly higher than that of immature DC, as analyzed by IL-2 and $IFN-\gamma$ production and proliferation of the responding T cells in the co-culture with allogeneic T cells. The antigen-presenting capacity of HemoHIN-treated DC was also increased by the co-culture with OVA-specific T cells (HS-1), as analyzed by IL-2 and $IFN-\gamma$ production and the proliferation. These results indicate that HemoHIM causes the maturation and ;Activation of DC, which may be a part of mechanisms of immunomodulation by HemoHIM.

Biosynthesis of Lactate-containing Polyhydroxyalkanoates in Recombinant Escherichia coli by Employing New CoA Transferases (재조합 대장균에서 새로운 코엔자임 에이 트랜스퍼레이즈를 이용한 젖산을 모노머로 함유한 폴리하이드록시알칸산 생산 연구)

  • Kim, You Jin;Chae, Cheol Gi;Kang, Kyoung Hee;Oh, Young Hoon;Joo, Jeong Chan;Song, Bong Keun;Lee, Sang Yup;Park, Si Jae
    • KSBB Journal
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    • v.31 no.1
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    • pp.27-32
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    • 2016
  • Several CoA transferases from Clostridium beijerinckii, C. perfringens and Klebsiella pneumoniae were examined for biosynthesis of lactate-containing polyhydroxyalkanoates (PHAs) in recombinant Escherichia coli XL1-Blue strain. The CB3819 gene and the CB4543 gene from C. beijerinckii, the pct gene from C. perfringens and the pct gene from K. pneumoniae, which encodes putative CoA transferase gene, respectively, was co-expressed with the Pseudomonas sp. MBEL 6-19 phaC1437 gene encoding engineered Pseudomonas sp. MBEL 6-19 PHA synthase 1 ($PhaC1_{Ps6-19}$) to examine its activity for the construction of key metabolic pathway to produce poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)]. The recombinant E. coli XL1-Blue expressing the phaC1437 gene and CB3819 gene synthesized poly(3-hydroxybutyrate) [P(3HB)] homopolymer to the P(3HB) content of 60.5 wt% when it was cultured in a chemically defined medium containing 20 g/L of glucose and 2 g/L of sodium 3-hydroxybutyrate. Expression of the phaC1437 gene and CB4543 gene in recombinant E. coli XL1-Blue also produced P(3HB) homopolymer to the P(3HB) content of 51.2 wt% in the same culture condition. Expression of the phaC1437 gene and the K. pneumoniae pct gene in recombinant E. coli XL1-Blue could not result in the production of PHAs in the same culture condition. However, the recombinant E. coli XL1-Blue expressing the phaC1437 gene and the C. perfringens gene could produce poly(3-hydroxybutyrate-co-lactate [P(86.4mol%3HB-co-13.7 mol%LA) up to the PHA content of 10.6 wt% in the same culture condition. Newly examined CoA transfereases in this study may be useful for the construction of engineered E. coli strains to produce PHA containing novel monomer such lactate.