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Effects of Lactococcus lactis subsp. lactis I2 with β-Glucooligosaccharides on Growth, Innate Immunity and Streptococcosis Resistance in Olive Flounder (Paralichthys olivaceus)

  • Hasan, Md Tawheed;Jang, Won Je;Tak, Jin Yeong;Lee, Bong-Joo;Kim, Kang Woong;Hur, Sang Woo;Han, Hyon-Sob;Kim, Bo-Seong;Huh, Min-Do;Kim, Shin-Kwon;Kong, In-Soo
    • Journal of Microbiology and Biotechnology
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    • v.28 no.9
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    • pp.1433-1442
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    • 2018
  • To identify and quantify the effects of a combination of dietary $1{\times}10^8CFU/g$ Lactococcus lactis subsp. lactis I2 ($LI_2$) and 0.1% ${\beta}$-glucooligosaccharides (BGO) on the growth and immunity of olive flounder (Paralichthys olivaceus), a feeding experiment was conducted. Flounder ($14{\pm}0.5g$) were divided into two groups and fed control and synbiotic feeds for 8 weeks. Investigations were carried out on growth and feed utilization, innate immunity, serum biochemical parameters, intestinal lactic acid bacterial (LAB) viability, microvillus length, and changes in the expression levels of genes encoding pro-inflammatory cytokines (tumor necrosis factor $[TNF]-{\alpha}$, interleukin $[IL]-1{\beta}$, and IL-6). Results demonstrated the synbiotic diet had significantly better (p < 0.05) responses in terms of weight gain and specific growth rate, three innate immune parameters (respiratory burst, serum lysozyme, and superoxide dismutase), intestinal LAB viability, and the relative $TNF-{\alpha}$ expression level (p < 0.05). Moreover, after challenge with Streptococcus iniae ($1{\times}10^8CFU/ml$), the synbiotically fed group exhibited significantly higher (p < 0.05) protection against streptococcosis, validating the observed changes in immune parameters and induction of the cytokine-encoding gene. Therefore, according to the results of the present study, synbiotic feed ($LI_2+BGO$) increased growth, modulated innate immune parameters and protected olive flounder against streptococcosis.

Building Light Weight CORBA Based Middleware for the CAN Bus Systems

  • Hong, Seongsoo
    • Transactions on Control, Automation and Systems Engineering
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    • v.3 no.3
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    • pp.181-189
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    • 2001
  • The software components of embedded control systems get extremely complex as they are designed into distributed systems get extremely complex as they are designed into distributed systems consisting of a large number of inexpensive microcontrollers interconnected by low-bandwidth real-time networks such as the controller area network (CAN). While recently emerging middleware technologies such as CORBA and DCOM address the complexity of distributed programming, they cannot be directly applied to distributed control system design due to their excessive resource demand and inadequate communication models. In this paper, we propose a CORBA-based middleware design for CAN-based distributed embedded control systems. Our design goal is to minimize its resource need and make it support group communication without losing the IDL (interface definition language) level compliance to the OMG standards. To achieve this, we develop a transport protocol on the CAN and a group communication scheme based on the well-known publisher/subscriber model. The protocol effectively realizes subject-based addressing and supports anonymous publisher/subscriber communication. We also customize the method invocation and message passing protocol, referred to as the general inter-ORB protocol (GIOP), of CORBA so that CORBA method invocations are efficiently serviced on a low-bandwidth network such as the CAN. This customization includes packed data encoding and variable-length integer encoding for compact representation of IDL data types. We have implemented our CORBA-based middleware on the mArx real-time operating system we have developed at Seoul National University. Our experiments clearly demonstrate that it is feasible to use CORBA in developing distributed embedded control systems possessing severe resource limitations. Our design clearly demonstrates that it is feasible to use a CORBA-based middleware in developing distributed embedded systems on real-time networks possessing severe resource limitations.

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Inter-frame vertex selection algorithm for lossy coding of shapes in video sequences (동영상에서의 모양 정보 부호화를 위한 정점 선택 알고리즘)

  • Suh, Jong-Yeul;Kim, Kyong-Joong;Kang, Moon-Gi
    • Journal of the Institute of Electronics Engineers of Korea SP
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    • v.37 no.4
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    • pp.35-45
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    • 2000
  • The vertex-based boundary encoding scheme is widely used in object-based video coding area and computer graphics due to its scalability with natural looking approximation. Existing single framebased vertex encoding algorithm is not efficient for temporally correlated video sequences because it does not remove temporal redundancy. In the proposed method, a vertex point is selected from not only the boundary points of the current frame but also the vertex points of the previous frame to remove temporal redundancy of shape information in video sequences. The problem of selecting optimal vertex points is modeled as finding shortest path in the directed acyclic graph with weight The boundary is approximated by a polygon which can be encoded with the smallest number of bits for maximum distortion. The temporal redundancy between two successive frames is efficiently removed with the proposed scheme, resulting in lower bit-rate than the conventional algorithms.

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Cloning of the dextranase gene(lsd11) from Lipomyces starkeyi and its expression in Pichia pastoris.

  • Park, Ji-Young;Kang, Hee-Kyoung;Jin, Xing-Ji;Ahn, Joon-Seob;Kim, Seung-Heuk;Kim, Do-Won;Kim, Do-Man
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.644-648
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    • 2005
  • Dextranase (${\alpha}$-1,6-D-glucan-6-glucanogydrolase:E.C. 3.2.1.11) catalyzes the hydrolysis of ${\alpha}$-(1.6) linkages of dextran. A lsd1 gene encoding an extracellular dextranase was isolated from the genomic DNA of L. starkeyi. The lsd11 gene is a synthetic dextranase (lsd1) after codon optimization for gene expression with Pichia pastoris system. A open reading frame of lsd11 gene was 1827 bp and it was inserted into the pPIC3.5K expression vector. The plasmid linearized by Sac I was integrated into the 5'AOX region of the chromosomal DNA of P. pastoris. The lsd11 gene fragment encoding a mature protein of 608 amino acids with a predicted molecular weight of 70 kDa, was expressed in the methylotrophic yeast P. pastoris by controling the alcohol oxidase-1 (AOX1) promoter. The recombinant lds11 was optimized by using the shake-flask expression and upscaled using fermentation technology. More than 9.8 mg/L of active dextranase was obtained after induction by methanol. The optimum pH of LSD11 was found to be 5.5 and the optimum temperature $28^{\circ}C$.

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Toxic Effects of Alumina Nanoparticles in Rat Cerebrums and Kidneys (산화알루미늄 나노물질이 랫드의 대뇌와 신장에 미치는 영향)

  • Jo, Eunhye;Seo, Gyun-Baek;Kim, Hyunmi;Choi, Kyunghee;Kwon, Jung-Taek;Kim, Philje;Eom, Igchun
    • Journal of Environmental Health Sciences
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    • v.42 no.1
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    • pp.27-33
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    • 2016
  • Objectives: Alumina nanoparticles ($Al_2O_3$, Al-NPs) are used for various purposes, including as coating agents and paint additives. Their potential toxicity has raised concern for human health. This study focuses on exploring the toxic effects on the brain and kidneys caused by Al-NPs exposure in rats. Methods: The animals were orally administered Al-NPs at 10, 50 and 100 mg/kg body weight for 28 days following OECD TG 407. To determine the targeted toxicity of Al-NPs, histopathological examination and gene expression analysis were conducted on the rats. Results: The Al-NPs treatment induced kidney tubular dilatation. In the rat cerebrums, the expression levels of 126 genes experienced two-fold or greater increases in response to Al-NPs, including other genes encoding proteins involved in cell differentiation, transcription and signal transduction. In the rat kidneys, the expression levels of 152 genes also showed two-fold or greater increases in response to Al-NPs, including other genes encoding proteins involved in apoptosis, transcription and signal transduction. Conclusion: These results suggest that exposure to Al-NPs influences cellular signal pathways of kidney and cerebrum, and it can be a toxic indicators of nanometrials.

Single-Chain Fv Fragment of Catalytic Antibody 4f4f with Glycosidase Activity: Design, Expression, and Purification

  • Jang, Chang-Hwan;Chung, Hyun-Ho;Yu, Jae-Hoon;Chang, Yung-Jin;Kim, Hyong-Bai;Paek, Se-Hwan;Shin, Dong-Hoon;Kim, Kyung-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.9 no.3
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    • pp.376-380
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    • 1999
  • Constructs, encoding a single-chain variable fragment of a catalytic antibody 4f4f (scFv-4f4f) with glycosidase activity, were made by combining the coding sequences for the heavy and light chain variable domains with a sequence encoding a linker (GGGGS). Using three different plasmid systems, single-chain antibodies were expressed separately in Escherichia coli, demonstrating significant differences in the expression level and amounts in soluble form of the recombinant protein. The protein expression from pET3a-scFv-4f4f was up to 20% of the total soluble proteins and, more importantly, the proteins were mostly found in a soluble form. An SDS-PAGE analysis of the purified single-chain proteins, yielding higher than 5mg from a 1-1 culture, showed a single band corresponding to its molecular weight of 29,100. A preliminary study shows that the expressed scFv-4f4f is catalytically active. The catalytic parameters for the hydrolysis of p-nitrophenyl-$\beta$-D-glucopyranoside by scFv-4f4f are being investigated.

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Characterization of the recombinant cellulase B from Thermotoga maritima (Thermotoga maritima 유래 내열성 cellulase B 융합단백질의 특성 규명)

  • Chung Ho Kim
    • Journal of Applied Biological Chemistry
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    • v.65 no.4
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    • pp.383-386
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    • 2022
  • A gene encoding thermostable cellulase B (TmCelB) was isolated from Thermotoga maritima. The open reading frame (ORF) of TmCelB gene was 825bp long which predicted to encode 274 amino acid residues with a molecular weight of 31,732 Da. The 17 amino acid residues from N-terminal of the TmCelB was known as signal peptides. To analyze the enzymatic activity and biochemical properties, the ORF of TmCelB gene excluding a putative signal sequence encoding 17 amino acids were introduced into the E. coli expression vector, pRSET-B, and overexpressed in E. coli BL21. The optimum temperature of recombinant TmCelB was around 95 ℃, and the optimum pH of recombinant TmCelB was around pH 4.5. The recombinant TmCelB was stable at temperature below 100 ℃.

Cloning and Characterization of Pseudomonas mucidolens Exoinulinase

  • Kwon, Young-Man;Kim, Hwa-Young;Choi, Yong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.10 no.2
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    • pp.238-243
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    • 2000
  • An exoinulinase (${\beta}-D-fructofuranosidase$) gene was cloned by chromosome walking along the upstream region of the endoinulinase gene of Pseudomonas mucidolens isolated from soil. the exoinulinase gene consisted of an ORF of 0,506 bp encoding a polypeptide of 501 amino acids with a deduced molecular weight of 55,000. The exoinulinase produced by the recombinant Escherichia coli $DH5{\alpha}$ strain was also purified to homogeneity as determined by SDS-PAGE and a zymogram. The molecular weight of the purified exoinulinase according to both SDS-PAGE and gel filtration matched the deduced molecular weight of the protein described above, thereby indicating that the native form of the exoinulinase was a monomer. The purified enzyme hydrolyzed activity value of 2.0. Furthermore, no inulo-oligomers were liberated from the inulin substrate in the enzymatic reaction mixtures incubated for 90 min at $55^{\circ}C$. Taken together, these results indicate that the purified ${\beta}-D-fructofuranosidase$ was an exoinulinase. The pH and temperature optima of the exoinulinase were pH 6.0 and $55^{\circ}C$, respectively. the enzymehad no apparent requirement for a cofactor, and its activity was completely inactivated by $Ag^{+},{\;}Hg^{2+},{\;}and{\;}Zn^{2+}$. Kinetic experiments gave $K_m,{\;}V_{max},{\;}and{\;}K_{cat}$ values for inulin of 11.5 mM, 18 nM/s, and $72{\;}s^{-1}$, respectively. the exoinulinase was fairly stable in broad pH conditions (pH 5-9), and at pH 6.0 it showed a residual activity of about 70% after 4 h incubation at $55^{\circ}C$.

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High Molecular Weight Glutenin Subunit in Common Wheat (Triticum aestivum L.) (밀의 고분자 글루테닌 단백질)

  • Lee, Jong-Yeol;Kim, Yeong-Tae;Kang, Chon-Sik;Lim, Sun-Hyung;Ha, Sun-Hwa;Ahn, Sang-Nag;Kim, Young-Mi
    • Korean Journal of Breeding Science
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    • v.43 no.5
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    • pp.479-489
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    • 2011
  • Gluten is the main functional component of wheat, and is the main source of the viscoelastic properties in a dough. One of the gluten group is glutenin, which is composed of high molecular weight (HMW) and low molecular weight (LMW) subunits. The HMW glutenin subunits (HMW-GS) have been shown to play a crucial role in determining the processing properties of the grain. They are encoded by the Glu-1 loci located on the long arms of homeologous group one chromosomes, with each locus comprising two genes encoding x- and y-type subunits. The presence of certain HMW subunits is positively correlated with good bread-making quality. The highly conserved N- and C- terminal contaning cystein residues which form interand intra-chain disulphide bonds. This inter chain disulphide bonds stabilize the glutenin polymers. In contrast, the repetitive domains that comprise the central part of the HMW-GS are responsible for the elastic properties due to extensive arrays of interchain hydrogen bonds. In this review, we discuss HMW-GS, HMW-GS structure and gluten elasticity, relationship between HMW-GS and bread wheat quality and genetic engineering of the HMW-GS.

Use of the Synthetic Gene Encoding the Truncated Human Rotavirus VP8* Protein in Escherichia coli for Production of Vaccine Candidates or Development of Diagnostic Antibodies (합성 유전자를 이용하여 Escherichia coli에서 백신 후보의 생산 혹은 진단용 항체의 개발을 위한 인간 rotavirus VP8* 부분 단백질의 발현)

  • Kim, Sang-Rae;Lee, Bheong-Uk
    • Journal of Life Science
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    • v.28 no.4
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    • pp.478-482
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    • 2018
  • Human rotavirus is a causative agent of acute diarrhea among children. The artificial gene encoding the truncated $VP8^*$ protein of human rotavirus A (serotype 1 strain WA) was synthesized according to the Escherichia coli codon preference. The synthetic $VP8^*$ gene also possessed the NdeI and HindIII restriction sites for the convenient in-frame cloning for translation and a 6-histidine tag at C-terminus for Ni+ affinity purification. Molecular weight of the truncated $VP8^*$ protein deduced from the nucleotide sequences of the artificial gene was a 19.7-kDa. This synthetic $VP8^*$ DNA fragment was inserted into the pT7-7 expression vector and transformed into E. coli BL21 (DE3). Transformants harboring the synthetic gene encoding the $VP8^*$ protein was induced by supplement of a final concentration of 0.05 mM ITPG at $20^{\circ}C$. Protein crude extract from the E. coli transformants was subjected to Western blotting with the mouse anti-rotavirus capsid antibody, showing ~20-kDa $VP8^*$ protein band. The truncated $VP8^*$ protein band was also observed by Western blotting using the rabbit polyclonal antibody serum made against the truncated $VP8^*$ protein. This study suggested that the synthetic gene could be used as an easy way to produce the antigenic vaccine candidate for control of virus-associated diseases or to develop antibodies for diagnostic purpose.