• 제목/요약/키워드: ${\delta}$-sequence

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Xylitol 생산에 최적화된 xylose reductase (GRE3)의 분비발현 시스템 (Secretory Expression System of Xylose Reductase (GRE3) for Optimal Production of Xylitol)

  • 정회명;김재운;김연희
    • 생명과학회지
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    • 제26권12호
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    • pp.1376-1382
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    • 2016
  • Xylitol은 식품 및 의료산업에서 이용가치가 높은 물질로, lignocellulosic biomass인 xylose의 환원으로부터 생산되며, 대부분 유전적으로 안전한 Saccharomyces cerevisiae 균주를 사용하여 생산되고 있다. 따라서 본 연구에서는 S. cerevisiae에서 xylitol을 효율적으로 생산하기 위해 xylose reductase를 code하는 GRE3 (YHT104W)유전자의 발현시스템을 구축하여, xylose reductase의 분비생산 및 xylitol 생산성을 조사하고자 하였다. 먼저 GRE3 유전자의 발현에 적합한 promoter의 선별을 위해 GAL10 promoter와 ADH1 promoter 하류에 각각 mating factor ${\alpha}$ ($MF{\alpha}$) signal sequence와 GRE3 유전자를 가진 pGMF-GRE3와 pAMF-GRE3 plasmid를 구축하였다. 각각의 plasmid는 S. cerevisiae $SEY2102{\Delta}trp1$균주에 형질전환되었고, $SEY2102{\Delta}trp1$/pGMF-GRE3와 $SEY2102{\Delta}trp1$/pAMF-GRE3 형질전환주가 선별되었다. 그 중 $SEY2102{\Delta}trp1$/pGMF-GRE3 균주에서 NADPH를 cofactor로 사용했을 때 0.34 unit/mg-protein의 xylose reductase 활성(total activity)을 보였고, ADH1 promoter를 가진 $SEY2102{\Delta}trp1$/pAMF-GRE3 균주에 비해 1.5배 높은 활성증가를 확인 할 수 있었다. 또한 두 균주에서 모두 91%의 분비효율을 보여 대부분의 재조합 xylose reductase가 세포 밖으로 효율적으로 발현 분비되었음을 알 수 있었다. $SEY2102{\Delta}trp1$/pGMF-GRE3 균주를 사용한 baffled flask 배양에서 xylitol 생산량을 조사해 본 결과, 20 g/l의 xylose로부터 12.1 g/l의 xylitol을 생산하였고, 소모된 xylose의 약 83%정도가 xylitol로 환원되었음을 알 수 있었다.

분화도 좋은 구강 편평상피세포암종에서 Dominant Negative p63 isoform의 발현 (EXPRESSION OF DOMINANT NEGATIVE p63 ISOFORM IN WELL-DIFFERENCIATED ORAL SQUAMOUS CELL CARCINOMA)

  • 김인수;김철환;김경욱
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제33권3호
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    • pp.191-198
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    • 2007
  • The p53 which is well known as tumor suppressor gene is located at 17p13. p53 is a sequence-specific DNA binding transcription factor that responds to certain cytotoxic stresses, such as DNA damage, by enhancing the transcription of genes that regulate cell-cycle progression as well as programmed cell death. The p63 gene that is located at 3q27-29, is recognized members of the p53 family, and responsible for the transcription of 6 isoforms. Three isoforms ($TAp63{\alpha}$, $TAp63{\beta}$, $TAp63{\gamma}$) contain an N-terminal transactivation (TA) domain and can induce apoptosis. The other 3 isoforms (${\Delta}Np63{\alpha}$, ${\Delta}Np63{\beta}$, ${\Delta}Np63{\gamma}$) lack the TA domain and may function in a dominant-negative fashion by inhibiting the transactivation functions of p53 and TAp63 proteins, and thus act as oncoproteins. A number of studies have investigated the role of p63 in human squamous cell carcinomas from different organs. Only a few studies have examined ${\Delta}Np63$ isoform in oral squamous cell carcinoma including normal epithelium. This study aimed to evaluate expression of ${\Delta}Np63$ isoform in human oral squamous cell carcinoma tissue and normal mucosa. The 3 cases of well differenciated oral squamous cell carcinoma specimen including adjacent normal mucosa were examined, and immunohistochemical study with monoclonal antibody(4A4) and tumor cell apoptosis analysis with Transmission Electon Microscopy were studied. And, RT-PCR analysis was done for expression of ${\Delta}Np63$ isoform. The results were as followed. 1. Normal gingiva showed the restricted p63 expression in basal cell layer. 2. Well differentiated squamous cell carcinoma showed mainly p63 expression in overall area of malignancy, especially in basal cell layer to adjacent stromal tissue. 3. Tumor cells around keratinized area with no p63 expression disclosed less micro-organelle in decreased size cytoplasm and severe chromatin margination with nuclear destruction that means apoptosis. 4. Comparison of mRNA expression of ${\Delta}Np63$ isoform by RT-PCR showed variable expression of ${\Delta}Np63$ isoform, but ${\Delta}Np63{\alpha}$ was most highly expressed in all 3 tumor specimen. From theses results, it should be suggested that ${\Delta}Np63$ isoform expression in well differentiated squamous cell carcinoma was closely related to tumor oncogenesis, expecially overexpression of ${\Delta}Np63{\alpha}$ is a most important factor in tumor genesis of oral squamous cell carcinoma.

인공발(Prosthetic Foot) 스프링용 유리섬유강화 적층재의 적층배향에 따른 층간분리거동 해석 (Analysis of Delamination Behavior on the Stacking Sequence of Prosthetic Foot Keel in Glass fiber Reinforced Laminates)

  • 송삼홍;김철웅
    • 대한기계학회논문집A
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    • 제27권4호
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    • pp.623-631
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    • 2003
  • It is considered that the application of advanced composite materials to the prostheses for the disables is important to improve their bio-mechanical performance. Particularly, energy storing foot prosthesis is mostly important to restore gait ability of the disables with low-extremity amputation since it could provide propulsion at terminal stance enhancing the disables ability to walk long distance even run and jump. Therefore, the energy storing spring of Prosthetic foot keel under cyclic bending moment use mainly of high strength glass fiber reinforced plastic. The main objective of this study was to evaluate the stacking sequence effect using the delamination growth rate(dA$_{D}$/dN) of energy storing spring in glass fiber reinforced plastic under cyclic bending moment. The test results indicated that the shape of delamination zone depends on stacking sequence in GFRP laminates. Delamination area(A$_{D}$) turns out that variable types with the contour increased non-linearly toward the damage zones.nes.

STATCOM에서 영상분 전류주입에 의한 셀간 전압평형화 제어의 향상 (Enhancement of Cell Voltage Balancing Control by Zero Sequence Current Injection in a Cascaded H-Bridge STATCOM)

  • 권병기;정승기;김태형
    • 전력전자학회논문지
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    • 제20권4호
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    • pp.321-329
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    • 2015
  • The static synchronous compensator (STATCOM) of cascaded H-bridge configuration accompanying multiple separate DC sides is inherently subject to the problem of uneven DC voltages. These DC voltages in one leg can be controlled by adjusting the AC-side output voltage of each cell inverter, which is proportional to the active power. However, when the phase current is extremely small, large AC-side voltage is required to generate the active power to balance the cell voltages. In this study, an alternative zero-sequence current injection method is proposed, which facilitates effective cell balancing controllers at no load, and has no effect on the power grid because the injected zero sequence current only flows within the STATCOM delta circuit. The performance of the proposed method is verified through simulation and experiments.

주기적 신호를 이용한 단일모드 광섬유의 상호 위상변조 해석 (Analysis of Cross-Phase Modulation using a periodic signal in a Single-Mode Fiber)

  • 이종형
    • 한국정보통신학회논문지
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    • 제19권12호
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    • pp.2963-2967
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    • 2015
  • 단일모드 광섬유를 사용한 WDM 시스템에서 CPM에 의한 성능저하를 이론적으로 분석하기 위해 펌프-프로브 구조에서 펌프신호가 주기적이라고 가정하였다. 주기적 펌프신호는 '0'과 '1'이 교대로 발생하는 경우를 모델링한 것으로 CPM에 의한 EOP를 이론적으로 예측할 수 있게 한다. 유도된 이론적 결과를 수치해석 결과와 비교하였으며, 그 결과 표준 단일모드 광섬유를 사용한 경우는 넓은 영역의 채널 간격 ${\Delta}f$에 대해 유도된 이론식이 수치해석의 결과와 잘 일치하고 분산천이 광섬유를 사용한 경우는 ${\Delta}f$>100GHz 에서 일치하였다. 분산천이 광섬유를 사용하여 ${\Delta}f$<100GHz 인 경우는 CPM뿐만 아니라 FWM에 의한 성능저하가 두드러지므로 이론적 결과와 수치 해석의 결과 사이의 편차가 증가한다. 펌브-프로브 구조가 광섬유의 비선형성중 CPM의 영향을 주로 분석하기 위한 것이므로 이 결과는 예상대로 이다.

Structure-Activity Relationships of 13- and 14-Membered Cyclic Partial Retro-Inverso Pentapeptides Related to Enkephalin

  • Hong, Nam-Joo
    • Bulletin of the Korean Chemical Society
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    • 제31권4호
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    • pp.874-880
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    • 2010
  • A series of 13- and 14-membered cyclic enkephalin analogs based on the moderately $\mu$ selective prototype compound Tyr-C[D-$A_2bu$-Gly-Phe-Leu] 8a were synthesized to investigate the structure-activity relationship. The modifications of sequence were mainly focused on two positions 3 and 5, critical for the selective recognition for $\mu$ and $\delta$ opioid receptors. The substitution of hydrophobic $Leu^5$ with hydrophilic $Asp^5$ derivatives led to Tyr-C[D-$A_2bu$-Gly-Phe-Asp(N-Me)] 7 and Tyr-C[D-Glu-Phe-gPhe-rAsp(O-Me)] 5, the peptides with a large affinity losses at both $\mu$ and $\delta$ receptors. The substitution of $Phe^3$ with $Gly^3$ led to Tyr-C[D-Glu-Gly-gPhe-rLeu] 3 and Tyr-C[D-Glu-Gly-gPhe-D-rLeu] 4, the peptides with large affinity losses at $\mu$ receptors, indicating the critical role of phenyl ring of $Phe^3$ for $\mu$ receptor affinities. One atom reduction of the ring size from 14-membered analogs Tyr-C[D-Glu-Phe-gPhe-(L and D)-rLeu] 6a, 6b to 13-membered analogs Tyr-C[D-Asp-Phe-gPhe-(L and D)-rLeu] 1, 2 reduced the affinity at both $\mu$ and $\delta$ receptors, but increased the potency in the nociceptive assay, indicating the ring constrain is attributed to high nociceptive potency of the analogs. For the influence of D- or L-chirality of $Leu^5$ on the receptor selectivity, regardless of chirality and ring size, all cyclic diastereomers displayed marked $\mu$ selectivity with low potencies at the $\delta$ receptor. The retro-inverso analogs display similar or more active at $\mu$ receptor, but less active at $\delta$ receptor than the parent analogs.

Molecular Cloning of a Delta-class Glutathione S-transferase Gene from Bombus ignitus

  • Park, Jong-Hwa;Yoon, Hyung-Joo;Gui, Zhong Zheng;Jin, Byung-Rae;Sohn, Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
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    • 제18권1호
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    • pp.28-32
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    • 2009
  • We describe here the cloning and characterization of a cDNA encoding the glutathione S-transferase (GST) from the bumblebee Bombus ignitus. The Delta-class B. ignitus GST (BiGSTD) gene spans 1668 bp and consists of four introns and five exons that encode 216 amino acid residues with a calculated molecular weight of approximately 24561 Da and a pI of 8.03. The N-terminal domain of BiGSTD has a conserved Ser residue, as well as conserved Lys, Pro, Glu, Ser and Tyr residues that are involved in the GSH-binding site of GST. The BiGSTD showed 60% protein sequence identity to the Bombyx mori GSTT1, 58% to Musca domestica GST, 57% to Drosophila melanogaster GST, and 55% to Anopheles gambiae GST1. BiGSTD was close to the insect-specific Delta class of GSTs in a phylogenetic tree. Northern blot analysis showed that BiGSTD is highly expressed in the fat body and midgut, and less so in the muscles of B. ignitus worker bees.

Identification and Heterologous Expression of a ${\Delta}4$-Fatty Acid Desaturase Gene from Isochrysis sphaerica

  • Guo, Bing;Jiang, Mulan;Wan, Xia;Gong, Yangmin;Liang, Zhuo;Hu, Chuanjiong
    • Journal of Microbiology and Biotechnology
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    • 제23권10호
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    • pp.1413-1421
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    • 2013
  • The marine microalga Isochrysis sphaerica is rich in the very-long-chain polyunsaturated fatty acids, including eicosapentaenoic acid (EPA, $C20:5{\omega}-3$) and docosahexaenoic acid (DHA, $C22:6{\omega}-3$) that are important to human health. Here, we report a functional characterization of a ${\Delta}4$-fatty acid desaturase gene (FAD4) from I. sphaerica. IsFAD4 contains a 1,284 bp open reading frame encoding a 427 amino acid polypeptide. The deduced amino sequence comprises three conserved histidine motifs and a cytochrome b5 domain at its N-terminus. Phylogenetic analysis indicated that IsFad4 formed a unique Isochrysis clade distinct from the counterparts of other eukaryotes. Heterologous expression of IsFAD4 in Pichia pastoris showed that IsFad4 was able to desaturate docosapentaenoic acid (DPA) to form DHA, and the rate of converting DPA to DHA was 79.8%. These results throw light on the potential industrial production of specific polyunsaturated fatty acids through IsFAD4 transgenic yeast or oil crops.

STRONG CONVERGENCE OF COMPOSITE IMPLICIT ITERATIVE PROCESS FOR A FINITE FAMILY OF NONEXPANSIVE MAPPINGS

  • Gu, Feng
    • East Asian mathematical journal
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    • 제24권1호
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    • pp.35-43
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    • 2008
  • Let E be a uniformly convex Banach space and K be a nonempty closed convex subset of E. Let ${\{T_i\}}^N_{i=1}$ be N nonexpansive self-mappings of K with $F\;=\;{\cap}^N_{i=1}F(T_i)\;{\neq}\;{\theta}$ (here $F(T_i)$ denotes the set of fixed points of $T_i$). Suppose that one of the mappings in ${\{T_i\}}^N_{i=1}$ is semi-compact. Let $\{{\alpha}_n\}\;{\subset}\;[{\delta},\;1-{\delta}]$ for some ${\delta}\;{\in}\;(0,\;1)$ and $\{{\beta}_n\}\;{\subset}\;[\tau,\;1]$ for some ${\tau}\;{\in}\;(0,\;1]$. For arbitrary $x_0\;{\in}\;K$, let the sequence {$x_n$} be defined iteratively by $\{{x_n\;=\;{\alpha}_nx_{n-1}\;+\;(1-{\alpha}_n)T_ny_n,\;\;\;\;\;\;\;\;\; \atop {y_n\;=\;{\beta}nx_{n-1}\;+\;(1-{\beta}_n)T_nx_n},\;{\forall}_n{\geq}1,}$, where $T_n\;=\;T_{n(modN)}$. Then {$x_n$} convergence strongly to a common fixed point of the mappings family ${\{T_i\}}^N_{i=1}$. The result presented in this paper generalized and improve the corresponding results of Chidume and Shahzad [C. E. Chidume, N. Shahzad, Strong convergence of an implicit iteration process for a finite family of nonexpansive mappings, Nonlinear Anal. 62(2005), 1149-1156] even in the case of ${\beta}_n\;{\equiv}\;1$ or N=1 are also new.

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Analysis for the function of the N-terminal repeat region of Bacillus polymyxa CFTase

  • Kim, Byoung-Woo;Park, Jung-Ha;Kim, Eun-Young;Kim, Kwang-Hyoun;Kwon, Hyoun-Ju
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XIII)
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    • pp.586-589
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    • 2003
  • Previously we reported the cloning and sequence analysis of a CFTase gene from Bacillus polymyxa. CFTase was divided into five distinct regions. In order to understand a role of the N-terminal repeat region on the function of CFTase from Bacillus polymyxa MGL21, deletion mutantCFTase ${\Delta}N$ was prepered. Recombinant protein was overproduced in E. coli as inclusion body, solubilized in bufer containing 8M urea, and refolded in the phosphate buffer. The molecular weight of the purified wild type CFTase and CFTase ${\Delta}N$ were 148kDa , 108kDa, respectively.

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