• Title/Summary/Keyword: signal sequence

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Purification and Characterization of a Novel Extracellular Thermostable Alkaline Protease from Streptomyces sp. M30

  • Xin, Yan;Sun, Zhibin;Chen, Qiongzhen;Wang, Jue;Wang, Yicheng;Luogong, Linfeng;Li, Shuhuan;Dong, Weiliang;Cui, Zhongli;Huang, Yan
    • Journal of Microbiology and Biotechnology
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    • v.25 no.11
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    • pp.1944-1953
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    • 2015
  • A novel alkaline protease from Streptomyces sp. M30, SapHM, was purified by ammonium sulfate precipitation, hydrophobic interaction chromatography, and DEAE-Sepharose chromatography, with a yield of 15.5% and a specific activity of 29,070 U/mg. Tryptic fragments of the purified SapHM were obtained by electrospray ionization quadrupole time-of-flight mass spectrometry. Nucleotide sequence analysis revealed that the gene sapHM contained 1,179 bp, corresponding to 392 amino acids with conserved Asp156, His187, and Ser339 residues of alkaline protease. The first 24 amino acid residues were predicted to be a signal peptide, and the molecular mass of the mature peptide was 37.1 kDa based on amino acid sequences and mass spectrometry. Pure SapHM was optimally active at 80℃ in 50 mM glycine-NaOH buffer (pH 9.0), and was broadly stable at 0-50℃ and pH 4.0-9.0. The protease relative activity was increased in the presence of Ni2+, Mn2+, and Cu2+ to 112%, 113%, and 147% of control, respectively. Pure SapHM was also activated by dimethylformamide, dimethyl sulfoxide, Tween 80, and urea. The activity of the purified enzyme was completely inhibited by phenylmethylsulfonyl fluoride, indicating that it is a serine-type protease. The Km and Vmax values were estimated to be 35.7 mg/ml, and 5 × 104 U/mg for casein. Substrate specificity analysis showed that SapH was active on casein, bovine serum albumin, and bovine serum fibrin.

Raising Visual Experience of Soccer Video for Mobile Viewers (이동형 단말기 사용자를 위한 축구경기 비디오의 시청경험 향상 방법)

  • Ahn, Il-Koo;Ko, Jae-Seung;Kim, Won-Jun;Kim, Chang-Ick
    • Journal of KIISE:Computing Practices and Letters
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    • v.13 no.3
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    • pp.165-178
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    • 2007
  • The recent progress in multimedia signal processing and transmission technologies has contributed to the extensive use of multimedia devices to watch sports games with small LCD panel. However, the most of video sequences are captured for normal viewing on standard TV or HDTV, for cost reasons, merely resized and delivered without additional editing. This may give the small-display-viewers uncomfortable experiences in understanding what is happening in a scene. For instance, in a soccer video sequence taken by a long-shot camera techniques, the tiny objects (e.g., soccer ball and players) may not be clearly viewed on the small LCD panel. Moreover, it is also difficult to recognize the contents of the scorebox which contains the elapsed time and scores. This renuires intelligent display technique to provide small-display-viewers with better experience. To this end, one of the key technologies is to determine region of interest (ROI) and display the magnified ROI on the screen, where ROI is a part of the scene that viewers pay more attention to than other regions. Examples include a region surrounding a ball in long-shot and a scorebox located in the comer of each frame. In this paper, we propose a scheme for raising viewing experiences of multimedia mobile device users. Instead of taking generic approaches utilizing visually salient features for extraction of ROI in a scene, we take domain-specific approach to exploit unique attributes of the soccer video. The proposed scheme consists of two modules: ROI determination and scorebox extraction. The experimental results show that the proposed scheme offers useful tools for intelligent video display on multimedia mobile devices.

Predicting Parturition Time through Ultrasonic Measurement of Posture Changing Rate in Crated Landrace Sows

  • Wang, J.S.;Wu, M.C.;Chang, H.L.;Young, M.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.5
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    • pp.682-692
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    • 2007
  • This study presents an automatic system to predict parturition time in the crated sows. The system relies on ultrasonic transducers mounted from above along the length of the crate. Using a 40 kHz time of flight (TOF) single envelope wave, the momentary distances between the sensors are measured. Therefore, the local momentary height of the sow and the momentary posture, i.e. standing posture (SDP), kneeling posture (KP), sitting posture (STP) and lateral lying posture (LLP) are determined. Crated sows change their postures from standing to lying and vice versa which follows a characteristic pattern. As parturition approaches, sows exhibit uneasiness, restlessness and the stand up sequence (SUS, the posture transition from LLP to SDP) rate increases because of labor pains. In time series, the SUS rate demonstrates a peak and it happens approximately 0-12 h before parturition. In this paper, the basic parturition threshold value method (BPTVM) and the same hour method (SHM) are proposed for predicting parturition, both of which are based on the SUS rate. The BPTVM mainly detects the peak of the SUS rate. As the SUS rate exceeds the threshold value, the parturition becomes predictable. Moreover, the SHM calculates the difference in the SUS rates between a particular time of day and the corresponding time of the preceding day. Compared to the BPTVM, the SHM can eliminate the circadian rhythm of the SUS rate influenced by feeding behavior. Using the SHM the parturition can be approximately predicted within hours. In an attempt to define the threshold parameters of predicting parturition, a data set with 32 sows of the SUS rate are used to estimate assumable predicting probability. The results show the assumable probability of the parturition prediction within 9 h is 96.9% for the SHM and 84.4% for the BPTVM. Moreover, the SHM can even reach a 75% probability of prediction within three hours of parturition. We conclude that the SHM is more accurate and is more useful for parturition time prediction. When parturition is detected, the proposed algorithm generates a warning signal which can inform human personnel to protect the mother and newborn piglets.

Characterization of a Multimodular Endo-β-1,4-Glucanase (Cel9K) from Paenibacillus sp. X4 with a Potential Additive for Saccharification

  • Lee, Jae Pil;Kim, Yoon A;Kim, Sung Kyum;Kim, Hoon
    • Journal of Microbiology and Biotechnology
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    • v.28 no.4
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    • pp.588-596
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    • 2018
  • An endo-${\beta}$-1,4-glucanase gene, cel9K, was cloned using the shot-gun method from Paenibacillus sp. X4, which was isolated from alpine soil. The gene was 2,994 bp in length, encoding a protein of 997 amino acid residues with a predicted signal peptide composed of 32 amino acid residues. Cel9K was a multimodular enzyme, and the molecular mass and theoretical pI of the mature Cel9K were 103.5 kDa and 4.81, respectively. Cel9K contains the GGxxDAGD, PHHR, GAxxGG, YxDDI, and EVxxDYN motifs found in most glycoside hydrolase family 9 (GH9) members. The protein sequence showed the highest similarity (88%) with the cellulase of Bacillus sp. BP23 in comparison with the enzymes with reported properties. The enzyme was purified by chromatography using HiTrap Q, CHT-II, and HiTrap Butyl HP. Using SDS-PAGE/activity staining, the molecular mass of Cel9K was estimated to be 93 kDa, which is a truncated form produced by the proteolytic cleavage of its C-terminus. Cel9K was optimally active at pH 5.5 and $50^{\circ}C$ and showed a half-life of 59.2 min at $50^{\circ}C$. The CMCase activity was increased to more than 150% in the presence of 2 mM $Na^+$, $K^+$, and $Ba^{2+}$, but decreased significantly to less than 50% by $Mn^{2+}$ and $Co^{2+}$. The addition of Cel9K to a commercial enzyme set (Celluclast 1.5L + Novozym 188) increased the saccharification of the pretreated reed and rice straw powders by 30.4% and 15.9%, respectively. The results suggest that Cel9K can be used to enhance the enzymatic conversion of lignocellulosic biomass to reducing sugars as an additive.

Three Non-Aspartate Amino Acid Mutations in the ComA Response Regulator Receiver Motif Severely Decrease Surfactin Production, Competence Development, and Spore Formation in Bacillus subtilis

  • Wang, Xiaoyu;Luo, Chuping;Liu, Youzhou;Nie, Yafeng;Liu, Yongfeng;Zhang, Rongsheng;Chen, Zhiyi
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.301-310
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    • 2010
  • Bacillus subtilis strains produce a broad spectrum of bioactive peptides. The lipopeptide surfactin belongs to one well-known class, which includes amphiphilic membrane-active biosurfactants and peptide antibiotics. Both the srfA promoter and the ComP-ComA signal transduction system are an important part of the factor that results in the production of surfactin. Bs-M49, obtained by means of low-energy ion implantation in wild-type Bs-916, produced significantly lower levels of surfactin, and had no obvious effects against R. solani. Occasionally, we found strain Bs-M49 decreased spore formation and the development of competence. Blast comparison of the sequences from Bs-916 and M49 indicate that there is no difference in the srfA operon promoter PsrfA, but there are differences in the coding sequence of the comA gene. These differences result in three missense mutations within the M49 ComA protein. RT-PCR analyses results showed that the expression levels of selected genes involved in competence and sporulation in both the wild-type Bs-916 and mutant M49 strains were significantly different. When we integrated the comA ORF into the chromosome of M49 at the amyE locus, M49 restored hemolytic activity and antifungal activity. Then, HPLC analyses results also showed the comA-complemented strain had a similar ability to produce surf actin with wild-type strain Bs-916. These data suggested that the mutation of three key amino acids in ComA greatly affected the biological activity of Bacillus subtilis. ComA protein 3D structure prediction and motif search prediction indicated that ComA has two obvious motifs common to response regulator proteins, which are the N-terminal response regulator receiver motif and the C-terminal helix-turn-helix motif. The three residues in the ComA N-terminal portion may be involved in phosphorylation activation mechanism. These structural prediction results implicate that three mutated residues in the ComA protein may play an important role in the formation of a salt-bridge to the phosphoryl group keeping active conformation to subsequent regulation of the expression of downstream genes.

Practical Considerations of Arterial Spin Labeling MRI for Measuring the Multi-slice Perfusion in the Human Brain (스핀 라벨링 자기공명영상을 이용한 사람 뇌에서의 뇌 관류영상의 현실적 문제점을 향상 시키는 방법 연구)

  • Jahng, Geon-Ho
    • Progress in Medical Physics
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    • v.18 no.1
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    • pp.35-41
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    • 2007
  • In this work practical considerations of a pulsed arterial spin labeling MRI are presented to reliable multi-slice perfusion measurements In the human brain. Three parameters were considered in this study. First, In order to improve slice profile and Inversion efficiency of a labeling pulse a high power Inversion pulse of adiabatic hyperbolic secant was designed. A $900^{\circ}$ rotation of the flip angle was provided to make a good slice profile and excellent Inversion efficiency. Second, to minimize contributions of a residual magnetization be4ween Interleaved scans of control and labeling we tested three different conditions which were applied 1) only saturation pulses, 2) only spotter gradients, and 3) combinations of saturation pulses and spotter gradients Applications of bo4h saturation pulses and spoiler gradients minimized the residual magnetization. Finally, to find a minimum gap between a tagged plane and an imaging plane we tested signal changes of the subtracted image between control and labeled Images with varying the gap. The optimum gap was about 20mm. In conclusion, In order to obtain high quality of perfusion Images In human brain It Is Important to use optimum parameters. Before routinely using In clinical studios, we recommend to make optimizations of sequence parameters.

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Performance of Radio Communication DS/CDMA System with Diversity Technique and BCH Coding under Impulsive Noise and Nakagami Fading (임펄스 잡음과 나카가미 페이딩이 존재할 때 다이버시티 기법과 오류정정 부호를 이용한 무선통신 DS/CDMA 시스템의 오율 특성)

  • 김지웅;강희조;이권현
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.10 no.4
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    • pp.539-549
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    • 1999
  • In this paper, the bit error rare (BER) performance of DS/CDMA DQPSK communication system in the presence of multi access interference, impulsive noise and Nakagami fading is investigated. The DS/CDMA DQPSK communication system adopts Maximum Ratio Combining (MRC) diversity reception and error correcting BCH code technique to enhance system performance. Using the derived error probability equation, the error rate performance of DS/CDMA DQPSK communication system has been evaluated and shown in figures to discuss as a function of impulsive index(A), Gaussian noise to impulsive noise power ratio($\Gamma$'), multi access interference(Κ), Nakagami fading parameter(m), the number of diversity branch (L), the number of error correction symbol (t), PN code sequence length(N) and $E_b/N_0$. The error performance of DS/CDMA-MDPSK signals improve by adopting MRC diversity and BCH(15,7) coding technique in the environment of impulsive noise plus Nagakami fading. From the results, we known that proposed system is affected by multi access interference, impulsive noise and Nakagami fading in radio communication system environment. Also, BER performance of DS/CDMA DQPSK communication system cam be improved increasing either the power of desired signal or the value of Gaussian noise to impulsive noise power ratio. And BCH(15,7) code technique is more effective to restrain the affection of multi access, interference, impulsive noise and Nakagami fading in DS/CDMA DQPSK communication system than MRC diversity reception technique.

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Performance Analysis of Multicarrier Code Selection CDMA System for PAPR Reduction in Multipath Fading Channel (PAPR을 줄이기 위한 Multicarrier Code Select CDMA시스템의 다중 경로 페이딩 채널에서 성능 분석)

  • Ryu Kwan Woong;Park Yong Wan;Hong Een Kee;Kim Myovng Jin
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.29 no.12A
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    • pp.1319-1332
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    • 2004
  • Multicarrier direct sequence code-division multiple access CDMA(MC DS-CDMA) is an attractive technique for achieving high data rate transmission even if the potentially large peak-to-average power ratio(PAPR) is an important factor for its application. On the other hand, code select CDMA(CS-CDMA) is an attractive technique with constant amplitude transmission of multicode signal irregardless of subchannels by introducing code selection method. In this paper we propose a new multiple access scheme based on the combination of MC DS-CDMA and CS-CDMA. Proposed scheme, which we called MC CS-CDMA, includes the sutclasses of MC DS-CDMA and CS-CDMA as special cases. The performance of this system is investigated for multipath Sequency selective fading channel and maximal ratio combining with rake receiver. In addition the PAPR of proposed system is compare with that of both MC BS-CDMA and CS-CDMA. Simulation results show that proposed system improves PAPR reduction than MC DS-CDMA at the expense of the complexity of receiver and the number of available non. Also, the numerical result shows that the proposed system is better performance than MC DS-CDMA due to the increasing processing gain and the number of time diversity gain.

Enhanced Production of hCTLA4Ig through Increased Permeability in Transgenic Rice Cell Cultures (형질전환 벼 현탁세포 배양에서 투과성 증진을 통한 hCTLA4Ig의 생산성 증대)

  • Choi, Hong-Yeol;Cheon, Su-Hwan;Kwon, Jun-Young;Lim, Jung-Ae;Park, Hye-Rim;Kim, Dong-Il
    • KSBB Journal
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    • v.31 no.4
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    • pp.277-283
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    • 2016
  • In this system, rice cells were genetically modified to express human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) using RAmy3D promoter induced by sugar depletion. Even though the target protein fused with signal sequence peptide, plant cell wall can be a barrier against secretion of recombinant proteins. Therefore, hCTLA4Ig can be trapped inside cell wall or remained in intracellular space. In this study, to enhance the secretion of hCTLA4Ig from cytoplasm and cell walls into the medium, permeabilizing agents, such as dimethyl sulfoxide (DMSO), Triton X-100 and Tween 20, were applied in transgenic rice cell cultures. When 0.5% (v/v) of DMSO was added in sugar-free medium, intracellullar hCTLA4Ig was increased, on the other hand, the secreted extracellular hCTLA4Ig was lower than that of control. DMSO did not give permeable effects on transgenic rice cell cultures. And Triton X-100 was toxic to rice cells and also did not give enhancing permeability of cells. When 0.05% (v/v) Tween 20 was added in rice cell cultures, however, intracellular hCTLA4Ig was lower than that of control cultures. And the maximum 44.76 mg/L hCTLA4Ig was produced for 10 days after induction, which was 1.4-fold increase compared to that of control cultures. Especially, Tween 20 at 0.05% (v/v) showed the positive effect on the secretion of hCTLA4Ig though the decrease of intracellular hCTLA4Ig. Also, Tween 20 as a non-toxic surfactant did not affect the cell growth, cell viability and protease activity. In conclusion, secretion of hCTLA4Ig could be increased by enhancing permeability of cells regardless of the cell growth, cell viability and protease activity.

Characterization of Pv92, a Novel Merozoite Surface Protein of Plasmodium vivax

  • Lee, Seong-Kyun;Wang, Bo;Han, Jin-Hee;Nyunt, Myat Htut;Muh, Fauzi;Chootong, Patchanee;Ha, Kwon-Soo;Park, Won Sun;Hong, Seok-Ho;Park, Jeong-Hyun;Han, Eun-Taek
    • Parasites, Hosts and Diseases
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    • v.54 no.4
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    • pp.385-391
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    • 2016
  • The discovery and understanding of antigenic proteins are essential for development of a vaccine against malaria. In Plasmodium falciparum, Pf92 have been characterized as a merozoite surface protein, and this protein is expressed at the late schizont stage, but no study of Pv92, the orthologue of Pf92 in P. vivax, has been reported. Thus, the protein structure of Pv92 was analyzed, and the gene sequence was aligned with that of other Plasmodium spp. using bioinformatics tools. The recombinant Pv92 protein was expressed and purified using bacterial expression system and used for immunization of mice to gain the polyclonal antibody and for evaluation of antigenicity by protein array. Also, the antibody against Pv92 was used for subcellular analysis by immunofluorescence assay. The Pv92 protein has a signal peptide and a sexual stage s48/45 domain, and the cysteine residues at the N-terminal of Pv92 were completely conserved. The N-terminal of Pv92 was successfully expressed as soluble form using a bacterial expression system. The antibody raised against Pv92 recognized the parasites and completely merged with PvMSP1-19, indicating that Pv92 was localized on the merozoite surface. Evaluation of the human humoral immune response to Pv92 indicated moderate antigenicity, with 65% sensitivity and 95% specificity by protein array. Taken together, the merozoite surface localization and antigenicity of Pv92 implicate that it might be involved in attachment and invasion of a merozoite to a new host cell or immune evasion during invasion process.